Developmental endocrinology of larval moulting in the tobacco hornworm, Manduca sexta.
A larval moult in the tobacco hornworm, Manduca sexta, involves an endocrine cascade that begins with the release of a cerebral peptide hormone, the prothoracicotropic hormone (PTTH). The release of PTTH is gated, occurs during the scotophase and appears to be developmentally cued. In fourth instar Manduca larvae, PTTH release into the haemolymph occurs as a single burst over a few hours during the head critical period, i.e. the time during which the head (brain) is needed for the initiation of the moult to the fifth (last) instar. Released PTTH activates the prothoracic glands (PGs), and within a few hours the cumulative effect of this event results in a dramatic increase in the haemolymph ecdysteroid titre, which then elicits the moult. An assessment of the capacity of the corpora allata (CA) to synthesize juvenile hormone (JH) in vitro indicates that the above sequence of endocrine events begins only when JH synthesis has reached a nadir for the instar. Since CA activity is an indirect measure of the haemolymph titre of the hormone, it is conceivable that the developmentally cued release of PTTH is permissively controlled by a decreasing haemolymph titre of JH. With the increase in the ecdysteroid titre which marks the end of this endocrine cascade, the CA again become active, presumably to cause the increase in the JH haemolymph titre which directs the larval moult. This investigation has thus established the temporal and quantitative dynamics of the PTTH-PG axis that drive larval moulting and provides insight into the interendocrine regulatory relationships that may exist between the ecdysteroids and JHs. These possible relationships and the role of the brain in their regulation are discussed.


Acquired adult flat foot secondary to posterior tibial-tendon pathology.
Nineteen patients with the clinical diagnosis of dysfunction of the posterior tibial tendon underwent surgical exploration. Four types of lesions were identified: avulsion of the tendon at the insertion (Group I), mid-substance rupture of the tendon (Group II), an in-continuity tear of the tendon (Group III), and no tendon tear, tenosynovitis only (Group IV). These conditions could not be separated preoperatively by clinical or radiographic means. The patients in Group I were treated by reinsertion of the tendon; in Group II, by flexor tendon transfer; and in Groups III and IV, by synovectomy. At follow-up, most patients in Group I reported no improvement, but the patients in Groups II, III, and IV showed both subjective and objective improvement. The signs and symptoms of dysfunction of the posterior tibial tendon are not specific for mid-substance ruptures of the tendon but also can occur with avulsions or synovitis, or perhaps from other, as yet undefined lesions.


Effect of serum from patients with essential hypertension on sodium transport in normal leucocytes.
This study has confirmed that serum from patients with essential hypertension inhibits sodium transport and elevates intracellular sodium in normal human leucocytes in vitro when compared with that of well matched normotensive controls. The magnitude of this effect was positively correlated with the diastolic blood pressure of the hypertensive patient. The degree of sodium transport inhibition conferred by the hypertensive's serum was correlated with the abnormal sodium transport in the hypertensive's own leucocytes. These results confirm the presence of a serum inhibitor of sodium transport in essential hypertension. The relationship between the inhibitory effect and severity of hypertension argues that it may be of mechanistic importance.


Use of CT, ED presentation and hospitalisations 12 months before and after a diagnosis of cancer in Western Australia: a population-based retrospective cohort study.
To examine the use of CT, emergency department (ED)-presentation and hospitalisation and in 12 months before and after a diagnosis of cancer. Population-based retrospective cohort study. West Australian linked administrative records at individual level. 104 009 adults newly diagnosed with cancer in 2004-2014. CT use, ED presentations, hospitalisations. As compared with the rates in the 12th month before diagnosis, the rate of CT scans started to increase from 2 months before diagnosis with an increase in both ED presentations and hospitalisation from 1 month before the diagnosis. These rates peaked in the month of diagnosis for CT scans (477 (95% CI 471 to 482) per 1000 patients), and for hospitalisations (910 (95% CI 902 to 919) per 1000 patients), and the month prior to diagnosis for ED (181 (95% CI 178 to 184) per 1000 patients) then rapidly reduced after diagnosis but remained high for the next 12 months. While the patterns of the health services used were similar between 2004 and 2014, the rate of the health services used during after diagnosis was higher in 2014 versus 2004 except for CT use in patients with lymphohaematopoietic cancer with a significant reduction. Our results showed an increase in demand for health services from 2 months before diagnosis of cancer. Increasing use of health services during and post cancer diagnosis may warrant further investigation to identify factors driving this change.


Stakeholder Perspectives on the Optimizing Patient Transfers, Impacting Medical Quality, and Improving Symptoms: Transforming Institutional Care (OPTIMISTIC) Project.
The need to reduce burdensome and costly hospitalizations of frail nursing home residents is well documented. The Optimizing Patient Transfers, Impacting Medical Quality, and Improving Symptoms: Transforming Institutional Care (OPTIMISTIC) project achieved this reduction through a multicomponent collaborative care model. We conducted an implementation-focused project evaluation to describe stakeholders' perspectives on (a) the most and least effective components of the intervention; (b) barriers to implementation; and (c) program features that promoted its adoption. Nineteen nursing homes participated in OPTIMISTIC. We conducted semistructured, qualitative interviews with 63 stakeholders: 23 nursing home staff and leaders, 4 primary care providers, 10 family members, and 26 OPTIMISTIC clinical staff. We used directed content analysis to analyze the data. We found universal endorsement of the value of in-depth advance care planning (ACP) discussions in reducing hospitalizations and improving care. Similarly, all stakeholder groups emphasized that nursing home access to specially trained, project registered nurses (RNs) and nurse practitioners (NPs) with time to focus on ACP, comprehensive resident assessment, and staff education was particularly valuable in identifying residents' goals for care. Challenges to implementation included inadequately trained facility staff and resistance to changing practice. In addition, the program sometimes failed to communicate its goals and activities clearly, leaving facilities uncertain about the OPTIMISTIC clinical staff's roles in the facilities. These findings are important for dissemination efforts related to the OPTIMISTIC care model and may be applicable to other innovations in nursing homes.


The Ross operation in children and young adults: a fifteen-year, single-institution experience.
The optimal operation for aortic valve disease in children and young adults remains controversial. The Ross operation offers avoidance of anticoagulation and the potential for growth but is technically demanding and creates double-valve disease. The goal of this study is to report our experience with the Ross operation and the need for reintervention at intermediate follow-up. A retrospective review of Ross operations in a single surgeon experience from 1992 to 2007 was conducted. All echocardiograms were reevaluated by a single cardiologist. The cohort included 54 patients with a mean age of 13.5 years (range 0.5 to 35 years). Pulmonary autograft implantation was accomplished using root replacement (n=43), root inclusion (n=9), and Dacron tube root replacement (n=2). Follow-up was available for 47 patients (87%) at a mean length of 6.4 years. There were no deaths. Kaplan-Meier estimates of freedom from explantation at 10 years were 100% for the autograft and 71% for the homograft. Autograft insufficiency at latest follow-up was trivial in 37 patients (82%), mild in 6 patients (13%), and moderate in 2 patients (4%). Reintervention for the homograft included balloon dilation in 3 children and conduit change in 5 children (all≤2 years old at initial operation). The Ross operation can be performed in children and adults with low mortality and can provide a durable result for the aortic valve with a low incidence of aortic insufficiency. The need for homograft replacement during follow-up in our series was primarily limited to children who were age 2 years or younger at initial operation.


Covalent Lectin Inhibition and Application in Bacterial Biofilm Imaging.
Biofilm formation by pathogenic bacteria is a hallmark of chronic infections. In many cases, lectins play key roles in establishing biofilms. The pathogen Pseudomonas aeruginosa often exhibiting various drug resistances employs its lectins LecA and LecB as virulence factors and biofilm building blocks. Therefore, inhibition of the function of these proteins is thought to have potential in developing "pathoblockers" preventing biofilm formation and virulence. A covalent lectin inhibitor specific to a carbohydrate binding site is described for the first time. Its application in the LecA-specific in vitro imaging of biofilms formed by P. aeruginosa is also reported.


Longitudinal Assessment Using Optical Coherence Tomography in Patients with Friedreich's Ataxia.
Ocular abnormalities occur frequently in Friedreich's ataxia (FRDA), although visual symptoms are not always reported. We evaluated a cohort of patients with FRDA to characterise the clinical phenotype and optic nerve findings as detected with optical coherence tomography (OCT). A total of 48 patients from 42 unrelated families were recruited. Mean age at onset was 13.8 years (range 4-40), mean disease duration 19.5 years (range 5-43), mean disease severity as quantified with the Scale for the Assessment and Rating of Ataxia 22/40 (range 4.5-38). All patients displayed variable ataxia and two-thirds had ocular abnormalities. Statistically significant thinning of average retinal nerve fibre layer (RNFL) and thinning in all but the temporal quadrant compared to controls was demonstrated on OCT. Significant RNFL and macular thinning was documented over time in 20 individuals. Disease severity and visual acuity were correlated with RNFL and macular thickness, but no association was found with disease duration. Our results highlight that FDRA is associated with subclinical optic neuropathy. This is the largest longitudinal study of OCT findings in FRDA to date, demonstrating progressive RNFL thickness decline, suggesting that RNFL thickness as measured by OCT has the potential to become a quantifiable biomarker for the evaluation of disease progression in FRDA.


Analysis of the gene start and gene end signals of human respiratory syncytial virus: quasi-templated initiation at position 1 of the encoded mRNA.
The gene start (GS) and gene end (GE) transcription signals of human respiratory syncytial virus (RSV) strain A2 were analyzed in helper-dependent monocistronic and dicistronic minireplicons which were complemented by a standard RSV strain. The GS signal, which is the start site for mRNA synthesis, is highly conserved for the first nine genes: 3'-CCCCGUUUA(U/C) (negative sense). This conserved version of the signal was analyzed by "saturation" mutagenesis, in which all 10 positions, as well as one downstream and one upstream position, were changed one at a time into each of the other three nucleotides. Most of the positions appear to contribute to the signal: positions 1, 3, 6, 7, and, in particular, 9 were the most sensitive, whereas position 5 was relatively insensitive. The effect of nucleotide substitution in the first position of the signal was examined further by cDNA cloning and sequence analysis of the residual mRNA which was produced. For the two mutants examined (1C to U, and 1C to A), the site of initiation was unchanged. However, the mRNAs were dimorphic with regard to the assignment of the 5'-terminal nucleotide: two-thirds contained the predicted mutant substitution, and one-third contained the parental assignment. Intracellular minigenome contained only the mutant assignment, indicating that the heterogeneity was at the level of transcription by the RSV polymerase. This suggests that the templated mutant assignment at position 1 can sometimes be overridden by an innate preference for the parental assignment, a phenomenon which we dubbed quasi-templated initiation. The GS signal of the L gene, encoding the 10th RSV mRNA, contains three differences (3'-CCCUGUUUUA) compared to the conserved version. It was shown to be equal in efficiency to the conserved version. This was unexpected, since the saturation mutagenesis described above indicated that U in place of A at position 9 should be highly inhibitory. Instead, the A at position 10 of the L GS signal was found to be critical for activity, indicating that an essential A residue indeed was present in both versions of the GS signal but that its spacing differed. The GE signal, which directs termination and polyadenylation, has more sequence diversity in nature than does the GS signal. The naturally occurring GE signals of strain A2 were compared by their individual incorporation into a dicistronic minigenome. They were similar in the ability to produce translatable mRNA except in the cases of NS1 and NS2, which were approximately 60% as efficient.


Genomic organization of the human TIMP-1 gene. Investigation of a causative role in the pathogenesis of X-linked retinitis pigmentosa 2.
To evaluate the role of TIMP-1 in inherited retinal degeneration. The genomic structure of the TIMP-1 gene was established and male patients with x-linked retinitis pigmentosa 2 from five families were screened for sequence alterations by direct sequencing in all exons, exon-intron boundaries, and the 5' upstream region of the gene. TIMP-1 appears to be expressed in the retina at low levels and consists of six exons spanning a genomic region of approximately 4.5 kb on Xp11.23. No disease-specific sequence alterations were identified. A site substitution in exon 5 was observed in samples from control subjects and patients, but it did not alter the amino acid sequence of the protein product. The results of this study exclude mutations in the TIMP-1 coding sequence, splice sites, and the 5' upstream region as a cause of retinal degeneration in x-linked retinitis pigmentosa 2. However, an as yet unidentified regulatory element that lies outside these intervals may be implicated. The role of this tightly regulated protein in the normal functioning of the retina has yet to be determined.


Different applications of isosbestic points, normalized spectra and dual wavelength as powerful tools for resolution of multicomponent mixtures with severely overlapping spectra.
Analysis of complex mixture containing three or more components represented a challenge for analysts. New smart spectrophotometric methods have been recently evolved with no limitation. A study of different novel and smart spectrophotometric techniques for resolution of severely overlapping spectra were presented in this work utilizing isosbestic points present in different absorption spectra, normalized spectra as a divisor and dual wavelengths. A quaternary mixture of drotaverine (DRO), caffeine (CAF), paracetamol (PCT) and para-aminophenol (PAP) was taken as an example for application of the proposed techniques without any separation steps. The adopted techniques adopted of successive and progressive steps manipulating zero /or ratio /or derivative spectra. The proposed techniques includes eight novel and simple methods namely direct spectrophotometry after applying derivative transformation (DT) via multiplying by a decoding spectrum, spectrum subtraction (SS), advanced absorbance subtraction (AAS), advanced amplitude modulation (AAM), simultaneous derivative ratio (S1DD), advanced ratio difference (ARD), induced ratio difference (IRD) and finally double divisor-ratio difference-dual wavelength (DD-RD-DW) methods. The proposed methods were assessed by analyzing synthetic mixtures of the studied drugs. They were also successfully applied to commercial pharmaceutical formulations without interference from other dosage form additives. The methods were validated according to the ICH guidelines, accuracy, precision, repeatability, were found to be within the acceptable limits. The proposed procedures are accurate, simple and reproducible and yet economic. They are also sensitive and selective and could be used for routine analysis of complex most of the binary, ternary and quaternary mixtures and even more complex mixtures.


Hodgkin's lymphoma presenting with an unusual horizontal Nystagmus and vertigo.
Neurological presentation of paraneoplastic syndromes is rare. They are often difficult to diagnose, especially when they precede the diagnosis of cancer. Hodgkin's lymphoma is associated with several paraneoplastic neurological syndromes such as cerebellar degeneration and dermato/polymyositis. Peripheral vestibular syndrome is uncommon presentation of these paraneoplastic syndromes. We report the case of a 52-year-old man with no prior medical history who presented to the otolaryngology clinic with vertigo precipitated by nystagmus, nausea and weight loss. Diagnostic workup revealed a nodular sclerosing variant of Hodgkin's lymphoma without paraneoplastic antibodies. The patient's symptoms resolved after institution of chemotherapy, radiotherapy and vestibular rehabilitation. Hodgkin's lymphoma has been reported to be associated with many paraneoplastic syndromes with neurological presentation in which peripheral vestibular syndrome is an uncommon one. Sometimes it can be the only presenting symptom of an unknown Hodgkin's lymphoma. This create a real diagnostic challenge for clinicians specially when paraneoplastic antibodies are negative. Chemotherapy and radiotherapy as treatment of the underlying disease is of a big benefit if started as early as possible.


Towards a needs-based design of the physical rehabilitation workforce in South Africa: trend analysis [1990-2017] and a 5-year forecasting for the most impactful health conditions based on global burden of disease estimates.
Rehabilitation can improve function in many people with chronic health conditions. It is important to consider priority conditions requiring rehabilitation, so it can be realistically positioned and costed in national health financing systems like South Africa (SA)'s proposed National Health Insurance (NHI). This paper describes temporal trends of top-ranked conditions on years lived with disability (YLDs) rates in SA, for which physical rehabilitation can ameliorate associated disability. This study is a systematic synthesis of publicly available Global Burden of Disease (GBD) 2017 estimates. The top 11 conditions contributing most to YLDs and for which evidence-based rehabilitation interventions exist were identified. Age-standardized rates per 100,000 and YLDs counts were extracted from 1990 to 2017. Significance of changes in temporal trends was determined using Mann-Kendall trend tests. Best-fit rates of yearly changes were calculated per condition, using GBD estimates (2012-2017), and extrapolated (by imposing the best-fit regression line onto results for each subsequent predicted year) as forecasts (2018-2022). Trends for YLDs counts per condition year (1990-2017) and forecasted values (2018-2022) showed an overall steady increase for all conditions, except HIV and respiratory conditions. YLDs counts almost doubled from 1990 to 2017, with a 17% predicted increase from 2017 to 2022. The proportionate contribution to YLDs counts reduced over time for all conditions, except HIV. Although age-standardized YLDs rates appear relatively stable over the analyzed periods for all conditions (except HIV, respiratory conditions and type 2 diabetes), trend changes in YLDs rates over 28 years were significant for all conditions, except neonatal (p = 0.855), hearing loss (p = 0.100) and musculoskeletal conditions (p = 0.300). Significant trend decreases were apparent for 4/9 conditions, implying that another 5/9 conditions showed trend increases over 28 years. Predicted all-age prevalence in 2022 suggests relatively large increases for cardiovascular disease and heart failure, and burns, while relative decreases are predicted for fractures and dislocations, stroke, and musculoskeletal conditions. Rehabilitation needs in SA are potentially massive and unmet, highlighting the need for innovative and context-specific rehabilitation that considers current local needs and projected changes. These findings should be considered when designing the NHI and other schemes in SA to ensure human and financial resources are deployed efficiently.


Ferritinophagy Mediated by Oxidative Stress-Driven Mitochondrial Damage Is Involved in the Polystyrene Nanoparticles-Induced Ferroptosis of Lung Injury.
Nanoplastics are a common type of contaminant in the air. However, no investigations have focused on the toxic mechanism of lung injury induced by nanoplastic exposure. In the present study, polystyrene nanoplastics (PS-NPs) caused ferroptosis in lung epithelial cells, which could be alleviated by ferrostatin-1, deferoxamine, and N-acetylcysteine. Further investigation found that PS-NPs disturbed mitochondrial structure and function and triggered autophagy. Mechanistically, oxidative stress-derived mitochondrial damage contributed to ferroptosis, and autophagy-dependent ferritinophagy was a pivotal intermediate link, resulting in ferritin degradation and iron ion release. Furthermore, inhibition of ferroptosis using ferrostatin-1 alleviated pulmonary and systemic toxicity to reverse the mouse lung injury induced by PS-NPs inhalation. Most importantly, the lung-on-a-chip was further used to clarify the role of ferroptosis in the PS-NPs-induced lung injury by visualizing the ferroptosis, oxidative stress, and alveolar-capillary barrier dysfunction at the organ level. In summary, our study indicated that ferroptosis was an important mechanism for nanoplastics-induced lung injury through different lung cells, mouse inhalation models, and three-dimensional-based lung-on-a-chip, providing an insightful reference for pulmonary toxicity assessment of nanoplastics.


Assessment of progress over the CASP experiments.
The quality of structure models produced in the CASP5 experiment has been compared with that in earlier CASPs. The most significant progress is in the fold recognition regime, where the development of meta-servers has allowed more accurate consensus models to be generated. In contrast to this, there is little evidence of progress in producing more accurate comparative models, particularly those based on sequence identities > 30%. For comparative models based on low-sequence identity and for fold recognition models, accuracy depends primarily on the fraction of the target structure that is similar to an available template, and the quality of the alignment. Overall, these results indicate that there are still no effective methods of improving model quality beyond that obtained by successfully copying a template structure. For models of proteins with previously unknown folds, there appears to be a pause in the previous consistent improvement. There is some evidence that more groups are producing top-quality models, however. Although specific progress between successive experiments is sometimes difficulty to identify, over the history of all the CASPs there has been steady, if sometimes slow, progress in all modeling regimes.


Semiochemical mediation of oviposition by the phlebotomine sandfly Lutzomyia longipalpis.
Extracts of rabbit food, hay and rabbit faeces elicited a positive oviposition response from gravid female Lutzomyia longipalpis sandflies (Diptera: Phlebotominae). Combined extract of rabbit food and oviposition pheromone had a synergistic effect on sandfly egg-laying, greatly increasing the number of eggs laid and resulting in a highly targeted response. Individually tubed flies, exposed to the combined extract, were shown to be 3.5 times more likely to survive oviposition and laid 2.5 times more eggs than control flies. A laboratory oviposition trap baited with the combined extract was tested in a cage and caught 31 (62%) of 50 gravid L. longipalpis over a 72h period.


Role of macrophage oxidative metabolism in resistance to vesicular stomatitis virus infection.
The role of oxygen metabolites in mediating virucidal activity was studied in two cloned macrophage-like cell lines. The parental cell line, J774.16, upon appropriate stimulation with either phorbol myristate acetate (PMA) or aggregated immunoglobulin, is induced to oxidize glucose via the hexose monophosphate shunt and produce O2- and H2O2. A variant derived from it, clone C3C, is defective in oxidative metabolism and cannot be stimulated to produce O2- or H2O2. Significant differences in yields of vesicular stomatitis virus (VSV) between stimulated clone 16 cells and unstimulated cells could be obtained only when low multiplicities were used for infection. Under the same conditions, PMA stimulation of the variant clone C3C produced no reduction in yields. The effect of PMA on virus yields in clone 16 was short-lived and dose dependent. PMA stimulation of either cell line had no effect on the number of infectious centers, suggesting that the antiviral effect was likely to be an extracellular, rather than an intracellular, one. Using glucose oxidase plus aglucose to generate H2O2 in solution, we observed that H2O2 alone is capable of killing limited amounts of VSV. The inactivation of VSV, both by H2O2 in solution and by activated clone 16 cells, could be inhibited by catalase. We conclude that intracellular resistance to VSV is primarily mediated through nonoxidative mechanisms, since activated macrophages can kill only a limited number of infectious virus particles extracellularly by means of secreted H2O2.


Improvement of European translational cancer research. Collaboration between comprehensive cancer centers.
Even though the increasing incidence of cancer is mainly a consequence of a population with a longer life span, part of this augmentation is related to the increasing prevalence of patients living with a chronic cancer disease. To fight the problem, improved preventive strategies are mandatory in combination with an innovative health care provision that is driven by research. To overcome the weakness of translational research the OECI is proposing a practical approach as part of a strategy foreseen by the EUROCAN+PLUS feasibility study, which was launched by the EC in order to identify mechanisms for the coordination of cancer research in Europe.


Development of SLE among "potential SLE" patients seen in consultation: long-term follow-up.
To identify factors associated with development of systemic lupus erythematosus (SLE) among patients evaluated at a tertiary care Lupus Center for potential SLE. We identified patients first seen at the Brigham and Women's Hospital Lupus Center between 1 January 1992 and 31 December 2012 and thought to have potential SLE by a board-certified rheumatologist. All had 1-3 SLE ACR criteria at initial visit and > 2 follow-up visits ≥ 3 months apart. We reviewed medical records through 15 May 2013 for: SLE signs and symptoms, autoimmune serologies, prescriptions and diagnoses by board-certified rheumatologists. Bivariable analyses and multivariable logistic regression models were used to identify independent predictors of developing SLE. Two hundred and sixty four patients met inclusion criteria. At initial visit, mean age was 39.2 (SD 12.4) years, 94% were female and 67% white. Mean number of SLE ACR criteria was 2.7 (SD 1.0) and 88% were antinuclear antibody (ANA) positive at initial consultation. Mean follow-up time was 6.3 (SD 4.3) years and 67% were prescribed hydroxychloroquine in follow-up. At most recent visit, 56 (21%) had been diagnosed with SLE; 47 (18%) were thought not to have SLE and 161 (61%) were still considered to have potential SLE. In multivariable regression models, oral ulcers (OR 2.40, 95% CI 1.03-5.58), anti-dsDNA (OR 2.59, 95% CI 1.25-5.35) and baseline proteinuria or cellular casts (OR 16.20, 95% CI 1.63-161.02) were independent predictors of developing SLE. The most common other final diagnoses included fibromyalgia, Sjögren's syndrome, mixed connective tissue disease and cutaneous lupus. Among patients with potential SLE at initial consultation, 21% were diagnosed with definite SLE within 6.3 years. Oral ulcers, anti-dsDNA and proteinuria or cellular casts were independent predictors of developing definite SLE. A better means of accurately identifying those who will develop SLE among those presenting with potential disease is necessary.


Analysis of Postnatal Mutant Phenotypes in Mice.
Viable homozygous mutant newborn mice may show effects of a mutation at any time during their development by exhibiting abnormal structure, function, or lethality. This overview guides the analysis of postnatal mice through gross anatomical assessment and the detection of visible phenotypes prior to weaning such as altered growth patterns, neurological problems, or abnormalities in movement or coordination. Advice on marking pups for identification purposes and providing adequate nutrition in the event of eating problems is given. After weaning and at the onset of puberty, different phenotypes may become manifest, including compromised growth and vigor and reproductive problems in males and/or females. Assessing infertility in each sex is addressed.


Lessons learned with laparoscopic management of complicated grades of acute appendicitis.
Laparoscopy has not been consolidated as the approach of first choice in the management of complicated appendicitis. Methodological flaws and absence of disease stratification criteria have been implicated in that less evidence. The objective is to study the safe and effectiveness of laparoscopy in the management of complicated appendicitis according to laparoscopic grading system. From January 2008 to January 2011, 154 consecutive patients who underwent a laparoscopic appendectomy for complicated appendicitis were evaluated in the prospective way. The patient's age ranged from 12 to 75 years old (31.7 ± 13.3) and 58.3% were male. Complicated appendicitis refers to gangrenous and/or perforated appendix and were graded as 3A (segmental necrosis), 3B (base necrosis), 4A (abscess), 4B (regional peritonitis) and 5 (diffuse peritonitis). The outcomes including operative time, infection complication, operative complications and conversion rate were chosen to evaluate the procedure. The grade 3A was the most frequent with 50 (32.4%) patients. The mean operative time was 69.4 ± 26.3 minutes. The grade 4A showed the highest mean operative time (80.1 ± 26.7 minutes). The wound and intra-abdominal infection rates were 2.6 and 4.6%, respectively. The base necrosis was the most important factor associated with the conversion (5.2%). The grades 4A and 5 were associated with greater possibility of intra-abdominal collection. There were no operative complications. The laparoscopic management of all complicated grades of acute appendicitis is safe and effective and should be the procedure of first choice. The laparoscopic grading system allows us to assess patients in the same disease stage.


Myasthenia gravis associated with wasp sting.
A clinical picture indistinguishable fro, but not therefore identical with, myasthenia gravis (ocular form) developed within 24 hours of a wasp sting. Because of the close temporal association of events, operative mechanisms may be either an immediate hypersensitivity reaction to some components of wasp venom or a direct toxic effect of these substances on acetylcholine synthesis, release, or degradation. Theoretical considerations from the literature and the lack of immunologic abnormalities in the patient suggest that the latter may be correct. Unique host conditions must have been present at the time of the sting, implying latent or subclinical myasthenia gravis.


Sleeping disturbances and predictor risk factors among type 2 diabetic mellitus patients.
Sleep disturbance is a major health issue among people with type 2 diabetes mellitus (T2DM). The Pittsburgh Sleep Quality Index (PSQI) has been the most widely used instrument to measure subjective sleep disturbance. The aim of this study was to determine the impact of sleeping factor structure of the PSQI as potential predictor for glycosylated hemoglobin A1c (HbA1C) among people living with T2DM in the Turkish community to facilitate its use in the clinical practice and research. This is a cross-sectional study and participants were between the age group of 25 and 65 years old who visited the diabetes and endocrinology department of Mega Medipol University Teaching Hospital, Istanbul. The PSQI was conducted on 871 patients with T2DM. Good sleep quality was defined as PSQI score <5. Multivariate logistic regression analysis was used to estimate the associated risk factors for the T2DM. The current study showed significant differences between male and female patients with respect to their age in years, body mass index (BMI) (kg/m2), physical activity, smoking habit, sheesha smoking, income, family history of metabolic syndrome, coronary heart disease (CHD), and PSQI. The results revealed significant differences between HbA1c ≤7 and females and HbA1c >7 T2DM patients with respect to gender, BMI (kg/m2), CHD, and PSQI. The study demonstrated significant differences between sleeping categories PSQI as good, average, and poor sleeping among T2DM patients with respect to age and gender. Meanwhile, significant differences were reported between sleeping categories among T2DM patients with respect to their: number of sleeping hours, wake-up time, sleeping time, HbA1c, fasting blood glucose, uric acid, and systolic and diastolic blood pressure. This study showed very strong statistically significant correlations between low HbA1c and poor sleep quality in patients with T2DM patients, including subjective sleep quality r = 0.763, sleep latency r = 0.327, sleep duration r = 0.472, habitual sleep efficiency r = 0.575, sleep disturbances r = 0.564, use of sleep medication r = 0.728, and daytime dysfunction r = 0.734. Multivariate stepwise logistic regression analysis revealed that Vitamin D (mmol/L) (P < 0.001), HbA1c (P < 0.001), duration of DM (P < 0.001), uric acid (mmol/L) (P < 0.001), systolic blood pressure mmHg (P = 0.006), diastolic blood pressure mmHg (P = 0.015), and BMI (P = 0.024) were considered at higher risk as the predictors for sleeping quality among T2DM patients. The results suggest a strong positive correlation between PSQI with HbA1c levels, systolic and diastolic blood pressure, age, BMI, among type 2 diabetic patients. This study ascertains that poor sleep quality may be due to elevated level of HbA1c, metabolic syndrome, diabetes, obesity, and/or hypertension.


Comparison of the diuretic effects of frusemide and the Karavi Panchaka Ayurveda decoction.
To investigate the diuretic, natriuretic and kaliuretic effects of the antihypertensive Ayurveda drug Karavi Panchaka decoction and compare it with the diuretic frusemide. An animal study using Sprague-Dawley rats. The volume of urine and the total sodium and potassium excreted in the urine by rats in response to orally fed Karavi Panchaka decoction were compared with rats fed with frusemide. Control experiments were done with rats receiving similar volumes of distilled water orally. The Ayurveda drug was prepared in accordance with the traditional method in the laboratory using medicinal plant specimens individually collected and identified. The volume of urine excreted during a 24-hour period following administration of the Ayurveda drug, frusemide or water was measured. The total sodium and potassium ion concentrations in the urine samples were determined using flame photometry. The Karavi Panchaka decoction clearly showed a statistically significant increase in urine excretion when compared with the control group that received only distilled water. The potassium ion excretion was significantly increased in the Karavi Panchaka decoction treated group when compared to the control group. This increase was statistically similar to that caused by frusemide. Neither drug had a significant effect on sodium ion excretion at the dosages used. Our results show that the Karavi Panchaka decoction significantly increases urine and potassium ion excretion in rats, but has no effect on sodium ion excretion at the dosage used. The effect of the Ayurveda drug on urine output as well as the sodium and potassium ion excretion is similar to that of frusemide administered at the dose used in our study.


Management of venous flap insufficiency: the role of low molecular weight heparin in salvage.
Flap loss resulting from venous insufficiency is a difficult issue, both with pedicled or free flaps. We examined the efficiency of low molecular weight heparin administration in various forms and dressing methods in venous insufficiency. Forty-five Sprague Dawley rats were included and inferior epigastric artery perforated island flaps were obtained from the abdominal skin of the animals. Nine animals were randomly allocated to each of five experimental groups: sham-control (Group 1), venous occlusion only (Group 2), occlusion with systemic enoxaparin (Group 3), occlusion with systemic enoxaparin + local enoxaparin infusion to the punctiform incisions (Group 4), and occlusion with systemic enoxaparin + local enoxaparin infusion to the punctiform incisions and enoxaparin-impregnated sponge dressing (Group 5). Group 5 had higher flap survival rates than the other groups. Groups 3 and 4 had numerically better vitality than the control group, but the difference was not significant. Low molecular weight heparin administration, with or without punctiform incisions, is not efficient in rescuing flaps with venous insufficiency. Specialised dressing methods are necessary to maintain bleeding.


Alcohol outlet policies and practices concerning sales to underage people.
Little is known about the relationship between alcohol outlet policies and practices in the United States and the likelihood that outlets will sell alcoholic beverages to persons under the legal drinking age. This study assessed the prevalence of such outlet-level policies and practices, and analyzed the relationships between outlet variables and outlets' actual propensity to sell alcoholic beverages to youth. The sample consisted of all outlets licensed for off-premise and a 40% random sample of outlets licensed for on-premise sale of alcoholic beverages in 15 small to medium-sized communities in Minnesota and Wisconsin (USA). Data on outlet characteristics, policies and practices were collected by a telephone survey of the owner or manager of each outlet. In addition, alcohol purchase attempts were conducted at each outlet by youthful-appearing study confederates. Generally, policies and practices that may reduce the likelihood of sales to youth were reported by a minority of outlets. Purchase success was found to be associated with a number of outlet characteristics, policies and practices. The implications of these findings for policy and future research are discussed.


Small-fenestra cochleostomy for MED-EL electrode.
Although the physical characteristics of cochlear implant electrode arrays vary substantially among manufacturers, many otolaryngologists have adopted a surgical cochleostomy technique that is similar for all of them. The objective of this study was to compare the insertion dynamics of the MED-EL Flex electrode using two different sized cochleostomies. Six temporal bones were prepared with either a small fenestra ( approximately 1.0 mm) or a large fenestra ( approximately 1.8 mm) cochleostomy and the electrodes inserted under video fluoroscopy. The resulting video images were analysed for depth of insertion as well as dynamic flexion of the electrode during insertion. The study demonstrated that small-fenestra cochleostomies have a 'pool cue' or 'billiard cue' effect, stabilizing the electrode cable and resulting in easier and deeper insertions. This is achieved by limiting the buckling of the electrode just inside the cochleostomy site.


Development, validation and application of a liquid chromatography-tandem mass spectrometry method for quantifying lurasidone in dried blood spot samples.
Background: A liquid chromatography-tandem mass spectrometry method for quantifying lurasidone in rat dried blood spot (DBS) samples was developed. Method: The analyte was extracted from DBSs using the liquid-liquid extraction method. Chromatographic separation was achieved using a C18, Phenomenex, 150 × 4.6 mm, 3.0 μm column. The mobile phase composed of methanol, acetonitrile and water (70:10:20 v/v/v) with 0.1% heptafluorobutyric acid performed well in terms of reducing the matrix effect and achieving shorter retention time. Result: The method was validated over a concentration range of 5.0 to 1200.0 ng/ml and supported by the evaluation of various validation parameters. Conclusion: This simple, sensitive and specific method proved to be a viable alternative sampling method with reduced logistics and blood sample storage expenses despite analytical challenges.


Ablation of neurons expressing melanin-concentrating hormone (MCH) in adult mice improves glucose tolerance independent of MCH signaling.
Melanin-concentrating hormone (MCH)-expressing neurons have been ascribed many roles based on studies of MCH-deficient mice. However, MCH neurons express other neurotransmitters, including GABA, nesfatin, and cocaine-amphetamine-regulated transcript. The importance of these other signaling molecules made by MCH neurons remains incompletely characterized. To determine the roles of MCH neurons in vivo, we targeted expression of the human diphtheria toxin receptor (DTR) to the gene for MCH (Pmch). Within 2 weeks of diphtheria toxin injection, heterozygous Pmch(DTR/+) mice lost 98% of their MCH neurons. These mice became lean but ate normally and were hyperactive, especially during a fast. They also responded abnormally to psychostimulants. For these phenotypes, ablation of MCH neurons recapitulated knock-out of MCH, so MCH appears to be the critical neuromodulator released by these neurons. In contrast, MCH-neuron-ablated mice showed improved glucose tolerance when compared with MCH-deficient mutant mice and wild-type mice. We conclude that MCH neurons regulate glucose tolerance through signaling molecules other than MCH.


Hageman factor-dependent pathways: mechanism of initiation and bradykinin formation.
The concentration of bradykinin in human plasma depends on its relative rates of formation and destruction. Bradykinin is destroyed by two enzymes: a plasma carboxypeptidase (anaphylatoxin inactivator) removes the COOH-terminal arginine to yield an inactive octapeptide, and a dipeptidase (identical to the angiotensin-converting enzyme) removes the COOH-terminal Phe-Arg to yield a fragment of seven amino acids that is further fragmented to an end product of five amino acids. Formation of bradykinin is initiated on binding of Hageman factor (HF) to certain negatively charged surfaces on which it autoactivates by an autodigestion mechanism. Initiation appears to depend on a trace of intrinsic activity present in HF that is at most 1/4000 that of activated HF (HFa); alternatively traces of circulating HFa could subserve the same function. HFa then converts coagulation factor XI to activated factor XI (XIa) and prekallikrein to kallikrein. Kallikrein then digests high-molecular-weight kininogen (HMW-kininogen) to form bradykinin. Prekallikrein and factor XI circulate bound to HMW-kininogen and surface binding of these complexes is mediated via this kininogen. In the absence of HMW-kininogen, activation of prekallikrein and factor XI is much diminished; thus HMW-kininogen has a cofactor function in kinin formation and coagulation. Once a trace of kallikrein is generated, a positive feedback reaction occurs in which kallikrein rapidly activates HF. This is much faster than the HF autoactivation rate; thus most HFa is formed by a kallikrein-dependent mechanism. HMW-kininogen is also therefore a cofactor for HF activation, but its effect on HF activation is indirect because it occurs via kallikrein formation. HFa can be further digested by kallikrein to form an active fragment (HFf), which is not surface bound and acts in the fluid phase. The activity of HFf on factor XI is minimal, but it is a potent prekallikrein activator and can therefore perpetuate fluid phase bradykinin formation until it is inactivated by the C1 inhibitor. In the absence of C1 inhibitor (hereditary angioedema) HFf may also interact with C1 and activate it enzymatically. The resultant augmented bradykinin formation and complement activation may account for the pathogenesis of the swelling characteristic of hereditary angioedema and the serologic changes observed during acute attacks.


M2-based influenza vaccines: recent advances and clinical potential.
Current influenza vaccines can prevent disease caused by influenza viruses but require annual administration and almost yearly reformulation. An attractive alternative approach would be to use a vaccine that provides broad and, ideally, lifelong protection against all influenza A and B virus strains. The extracellular domain of matrix protein 2 (M2e) of influenza A viruses is conserved and thus fits well in such a broadly protective vaccine. Areas covered: Recent advances in M2e vaccine design, the mode of action of M2e-based immunity and clinical progress of M2-based influenza vaccines. Expert commentary: Many M2e vaccine have been successfully tested for efficacy against a panel of divergent influenza viruses in animal models. More recently, clinical studies have been conducted with M2e vaccine candidates, which demonstrated their safety and immunogenicity in humans. Efficacy studies in humans are still needed to provide evidence that an M2e-based vaccine can protect against human influenza.


The psychometric function and reaction times of automated perimetry in normal and abnormal areas of the visual field in patients with glaucoma.
To study the relationship of reaction time to the psychometric function in normal subjects, normal sensitivity test locations in patients with glaucoma, and test locations with 10 to 20 dB loss in patients with glaucoma. The authors tested 10 patients with glaucoma and 10 age-matched normal volunteers with the Humphrey perimeter, first with program 24-2 and then with the method of constant stimuli to generate frequency of seeing curves. At two widely separated visual field locations on the program 24-2 grid, they presented stimuli in 2-dB intervals, 10 dB either side of the program 24-2 threshold, at 0 dB and 60 dB (15 repetitions per intensity). For the patients with glaucoma, they chose a visual field location with normal sensitivity and a location in an area of 10 to 20 dB loss. Analysis of variance with post hoc t-tests showed that reaction time (RT) at the 0-dB intensity was prolonged by approximately 90 msec in the abnormal sensitivity test location of patients with glaucoma compared to the control and the glaucoma normal sensitivity groups (P<0.0001). However, this difference was accounted for by only 4 of the 10 patients with glaucoma, reaching 100% of stimuli seen with the brightest stimulus at the moderately damaged test location. Reaction time at the frequency of seeing 50% estimated threshold showed no significant differences among the groups. Prolongation of RT from the 0-dB value was analyzed as a function of increasing attenuation of stimulus intensity. The results fit the equation RT = a + b(Intensity3) for all groups. There is no significant difference in RT between normal subjects and patients with glaucoma either at threshold or to suprathreshold stimuli. Reaction time increases after a power function with increasing attenuation of stimulus intensity up to the threshold.


[New conditions for evaluating static compliance].
Static compliance (Cst), time course and clinical and diagnostic significance of relationship between Cst and Vinsp were studied in 97 patients with congenital heart diseases aged over 3 years after open-heart surgery for correction of the defect during uneventful immediate postoperative period. Three groups of patients were distinguished: 1) patients operated on for Fallot's tetralogy without a previous aortopulmonary anastomosis (TF-1); 2) patients with Fallot's tetralogy with a previous systemic pulmonary anastomosis (TF-2); 3) patients operated on for atrioventricular septal defects and incomplete atrioventricular communication. Age subgroups of 3-5, 6-8, 9-11, and 12-15 years were singled out in each group. The proposed modified method for measuring Cst in children aged over 3 years operated on for congenital heart disease more accurately and reliably characterized the status of lung parenchyma. As a result, the correlation (r) between Cst and Vinsp in all nosological groups and age subgroups was much higher than was previously reported.


Quaternary ammonium compounds as structural probes of single batrachotoxin-activated Na+ channels.
Quaternary ammonium (QA) blockers are well-known structural probes for studying the permeation pathway of voltage-gated K+ channels. In this study we have examined the effects of a series of n-alkyl-trimethylammonium compounds (Cn-QA) on batrachotoxin (BTX)-activated Na+ channels from skeletal muscle incorporated into planar lipid bilayers. We found that these amphipathic QA compounds (Cn-QA where n = 10-18) block single Na+ channels preferentially from the internal side with equilibrium dissociation constants (KD) in the submicromolar to micromolar range. External application of amphipathic QA compounds is far less effective, by a factor of greater than 200. The block can be described by a QA molecule binding to a single site in the Na+ channel permeation pathway. QA binding affinity is dependent on transmembrane voltage with an effective valence (delta) of approximately 0.5. QA dwell times (given as mean closed times, tau c) increase as a function of n-alkyl chain length, ranging from approximately 13 ms for C10-QA to 500 ms for C18-QA at +50 mV. The results imply that there is a large hydrophobic region within the Na+ channel pore which accepts up to 18 methylene groups of the Cn-QA cation. This hydrophobic domain may be of clinical significance since it also interacts with local anesthetics such as cocaine and mepivacaine. Finally, like BTX-activated Na+ channels in bilayers, unmodified Na+ channels in GH3 cells are also susceptible to QA block. Amphipathic QA cations elicit both tonic and use-dependent inhibitions of normal Na+ currents in a manner similar to that of local anesthetic cocaine. We conclude that amphipathic QA compounds are valuable structural probes to study the permeation pathway of both normal and BTX-activated Na+ channels.


Acellular dermal allograft for sellar repair after transsphenoidal approach to pituitary adenomas.
Objectives Our practice has transitioned from using fat autograft to acellular dermal matrix (AlloDerm, LifeCell Corp, Woodlands, Texas, USA). We present the largest series to our knowledge of AlloDerm for sellar floor repair after transsphenoidal approach to pituitary adenoma and compare rates of postoperative cerebrospinal fluid (CSF) leak with an earlier cohort of patients whose CSF leaks were repaired with fat autograft. Design This is a retrospective cohort study comparing sellar repair with fat autograft versus inlay Alloderm between the years 2003 and 2012. The primary end point was postoperative CSF leak. Results A total of 429 patients (368 primary; 83 revision operations) without intraoperative lumbar drainage were included. A total of 18 postoperative CSF leaks were observed (3.9%). Intraoperative CSF leak occurred in 160 cases (35.5%). Among this subset of patients with intraoperative CSF leak, 95 underwent repair with AlloDerm and 46 underwent repair with fat autograft, with postoperative CSF leak rates of 8.4% and 15.2%, respectively (p = 0.34, chi-square test); 19 patients underwent repair with other techniques or no repair at all, with postoperative leak rate of 0%. Conclusions AlloDerm is an effective alternative to fat autograft in cases of low-flow CSF leak following transsphenoidal resection of pituitary adenoma.


Rationale for topical antibiotics.
Topical antibiotics are under attack by critics who claim that no proof of efficacy exists and that systemic administration is far more effective. In this paper, we review the existing evidence and conclude that topical antibiotics have a place in the prevention and treatment of superficial skin infections. Our own experimental studies showed that topical antibiotics promptly reduce the density of Staphylococcus aureus in secondarily infected chronic dermatoses, associated with clinical improvement. In addition, topical antibiotics were shown to be highly effective in preventing and suppressing experimental S. aureus infections in human volunteers.


Association between Altitude and the Framingham Risk Score: A Cross-Sectional Study in the Peruvian Adult Population.
To determine the association between altitude and the Framingham risk score in the Peruvian population, we performed a cross-sectional analytical study of data collected by the 2017-2018 Food and Nutrition Surveillance by Life Stages survey. The outcome of this study was the Framingham 10-year cardiovascular disease event risk prediction, which is composed of six modifiable and non-modifiable coronary risk factors. A generalized linear model (GLM) of the gamma family and log link function was used to report the crude and adjusted β coefficients. Several sensitivity analyses were performed to assess the association of interest. Data from a total of 833 surveyed participants were included. After adjusting for educational level, poverty level, alcohol consumption, physical activity level, the presence of any limitation, obesity, and area of residence, it was observed that altitude ≥ 2500 m above sea level (β = -0.42 [95% CI: -0.69 to -0.16]) was negatively and significantly associated with a decrease in the Framingham 10-year risk score. High altitude was significantly and negatively associated with Framingham 10-year risk scores. Our results will allow prevention strategies considering modifiable risk factors to avoid the development of cardiovascular diseases, especially in people living at low altitudes.


Male goldfish reproductive behaviour and physiology are severely affected by exogenous exposure to 17beta-estradiol.
Mature male goldfish were exposed to different concentrations of the natural hormone 17beta-estradiol (E2). Two methods of exposure were employed, via ingestion at 0, 1, 10 and 100 microg/g food and via the water at 0, 1 and 10 microg/l. The fish were exposed for 24-28 days during the spawning period. The males were then paired with an artificially induced, spawning female and their sexual behaviour was observed during a 15 min period. The physiological status of the fish was also examined with respect to GSI, presence of milt and spawning tubercles and the blood plasma concentration of E2. Despite the relatively short exposure period, exposure to physiological levels of E2 was shown to severely affect the male goldfish reproductive behaviour and physiology. In conclusion, the results from this study and the ability to interpret the effects on this well-studied species, show that the effects of E2, and possibly other estrogenic EDCs, have severe effects at several vital levels of male goldfish reproduction. The results also suggests that the hormone E2 can act as an endocrine disrupting chemical (EDC) in the environment.


Noise Outcomes in Servicemembers Epidemiology (NOISE) Study: Design, Methods, and Baseline Results.
Military Service members and Veterans commonly report hearing loss and tinnitus, both of which can result in significant disability. During military service, Service members are exposed to many different types of loud noise, which is strongly associated with hearing loss and tinnitus. Other military-related exposures, such as chemicals and traumatic brain injury (TBI), are also linked with auditory problems. The purpose of the "Noise Outcomes in Servicemembers Epidemiology" (NOISE) study is to gather information from Active-Duty Service members and recently separated Veterans about their military and nonmilitary noise exposures, other relevant military and nonmilitary exposures, and potential outcomes of these exposures including tinnitus, hearing loss, and other hearing-related health concerns. The NOISE study assesses lifetime noise exposures, chemical and blast exposures, TBI, physical and psychiatric comorbidities, and other military and nonmilitary exposures and outcomes that can affect auditory function. Participants undergo comprehensive in-person audiologic examinations; those who experience tinnitus undergo a complete tinnitus assessment. Exposures and select outcomes are reassessed annually by mail, and the comprehensive in-person assessment is completed every 5 years. This report presents descriptive, baseline data obtained from the first 690 participants enrolled between 2014 and 2018. Some notable findings from this analysis include: (1) the prevalence of hearing loss in the sample was 8% for low frequencies (0.25 to 2 kHz), 20% for high frequencies (3 to 8 kHz), and 39% for extended high frequencies (9 to 16 kHz); (2) the prevalence of tinnitus was 53%; (3) the prevalence of both hearing loss and tinnitus was higher among those with higher age, more years of military service, greater degree of noise exposure, and exposures to blasts and/or TBI in the military; and (4) tinnitus was most prevalent among participants who serve/served in the Army relative to the other military branches. The NOISE study is acquiring comprehensive data on military-related auditory dysfunction. It is the first of its kind to enroll active Service members and recently separated Veterans into a longitudinal study to examine the etiology and outcomes of tinnitus and hearing loss in this population. Although these data do not necessarily represent the entire military and Veteran populations, ongoing enrollment is focused on increasing generalizability and will also provide the statistical power to conduct multivariable analyses. This will allow us to examine longitudinal associations of interest while controlling for potential confounders and other possible sources of error. These data will provide critical knowledge to refine future military hearing conservation efforts and inform efforts to develop future treatments.


Mouse models of immune dysfunction: their neuroanatomical differences reflect their anxiety-behavioural phenotype.
Extensive evidence supports the role of the immune system in modulating brain function and behaviour. However, past studies have revealed striking heterogeneity in behavioural phenotypes produced from immune system dysfunction. Using magnetic resonance imaging, we studied the neuroanatomical differences among 11 distinct genetically modified mouse lines (n = 371), each deficient in a different element of the immune system. We found a significant and heterogeneous effect of immune dysfunction on the brains of both male and female mice. However, by imaging the whole brain and using Bayesian hierarchical modelling, we were able to identify patterns within the heterogeneous phenotype. Certain structures-such as the corpus callosum, midbrain, and thalamus-were more likely to be affected by immune dysfunction. A notable brain-behaviour relationship was identified with neuroanatomy endophenotypes across mouse models clustering according to anxiety-like behaviour phenotypes reported in literature, such as altered volume in brains regions associated with promoting fear response (e.g., the lateral septum and cerebellum). Interestingly, genes with preferential spatial expression in the most commonly affected regions are also associated with multiple sclerosis and other immune-mediated diseases. In total, our data suggest that the immune system modulates anxiety behaviour through well-established brain networks.


Herb-drug interaction of Epimedium sagittatum (Sieb. et Zucc.) maxim extract on the pharmacokinetics of sildenafil in rats.
Epimedium sagittatum (Sieb. et Zucc.) Maxim is one of the herbs used to treat erectile dysfunction in Traditional Chinese Medicine. Sildenafil is a phosphodiesterase 5 inhibitor used to treat erectile dysfunction in Western Medicine. This study evaluates the herbal-drug interaction of Epimedium sagittatum extract on the pharmacokinetics of sildenafil in rats by ultra-performance liquid chromatography. The rat plasma was sampled from each anesthetized rat after pretreatment with 3-days Epimedium sagittatum extract (1/2 g/kg/day) and intravenous injection with sildenafil (10/30 mg/kg). The pharmacokinetic data demonstrate that the area under the concentration-time curve (AUC) of sildenafil (10 mg/kg) was significantly decreased in groups that received a high dose of Epimedium sagittatum extract. In conclusion, the study demonstrates that there was significant herb-drug interaction of Epimedium sagittatum extract on the pharmacokinetics of sildenafil at low and high daily doses, suggesting co-administration use of Epimedium sagittatum extract and sildenafil in clinical practice should be prevented due to possible herb-drug interactions.


Molecular view on protein sorting into liquid-ordered membrane domains mediated by gangliosides and lipid anchors.
We present results from coarse grain molecular dynamics simulations of mixed model membranes consisting of saturated and unsaturated lipids together with cholesterol, in which lipid-anchored membrane proteins are embedded. The membrane proteins studied are the peripherally bound H-Ras, N-Ras, and Hedgehog, and the transmembrane peptides WALP and LAT. We provide a molecular view on how the presence and nature of these lipid anchors affects partitioning of the proteins between liquid-ordered and liquid-disordered domains. In addition, we probed the role of the ganglioside lipid GM1 on the protein sorting, showing formation of GM1-protein nano-domains that act as shuttles between the differently ordered membrane regions.


Centrally infused atrial natriuretic polypeptide attenuates exaggerated salt appetite in spontaneously hypertensive rats.
We have previously shown that atrial natriuretic polypeptide is present in the brain with the highest concentration in the hypothalamus and septum and that intracerebroventricular injection of atrial natriuretic polypeptide inhibits water drinking induced by centrally injected angiotensin II or 24-hour water deprivation in rats. To study further the role of brain atrial natriuretic polypeptide in the control of water and electrolyte balance, the effect of chronic intracerebroventricular infusion of atrial natriuretic polypeptide on salt appetite in spontaneously hypertensive rats and normotensive Wistar-Kyoto rats was examined with a free-choice, two-bottle preference test. The intracerebroventricular infusion of 100 ng/hour and 500 ng/hour of alpha-human atrial natriuretic polypeptide preferentially suppressed the intake of 0.30 M NaCl solution and attenuated the elevated preference for the hypertonic saline in spontaneously hypertensive rats while centrally infused alpha-human atrial natriuretic polypeptide had no significant effects on drinking behavior in Wistar-Kyoto rats. Blood pressure did not change significantly throughout the experiment in either rat strain. It is concluded that the exaggerated salt appetite in spontaneously hypertensive rats is blunted by centrally administered atrial natriuretic polypeptide. Such an effect of atrial natriuretic polypeptide along with its antidipsogenic effect suggests that brain atrial natriuretic polypeptide plays a role in water and electrolyte homeostasis and in blood pressure control.


The landscape of tyrosine kinase inhibitors in sarcomas: looking beyond pazopanib.
Introduction: Tyrosine kinases are key mediators of intracellular signaling cascades and aberrations in these proteins have been implicated in driving oncogenesis through the dysregulation of fundamental cellular processes including proliferation, migration, and apoptosis. As such, targeting these proteins with small molecule tyrosine kinase inhibitors (TKI) has led to significant advances in the treatment of a number of cancer types.Areas covered: Soft tissue sarcomas (STS) are a heterogeneous and challenging group of rare cancers to treat, but the approval of the TKI pazopanib for the treatment of advanced STS demonstrates that this class of drugs may have broad utility against a range of different sarcoma histological subtypes. Since the approval of pazopanib, a number of other TKIs have entered clinical trials to evaluate whether their activity in STS matches the promising results seen in other solid tumors. In this article, we review the emerging role of TKIs in the evolving landscape of sarcoma treatment.Expert opinion: As our biological understanding of response and resistance of STS to TKIs advances, we anticipate that patient management will move away from a 'one size fits all' paradigm toward personalized, multi-line, and patient-specific treatment regimens where patients are treated according to the underlying biology and genetics of their specific disease.


Tactile perception of randomly rough surfaces.
Most everyday surfaces are randomly rough and self-similar on sufficiently small scales. We investigated the tactile perception of randomly rough surfaces using 3D-printed samples, where the topographic structure and the statistical properties of scale-dependent roughness were varied independently. We found that the tactile perception of similarity between surfaces was dominated by the statistical micro-scale roughness rather than by their topographic resemblance. Participants were able to notice differences in the Hurst roughness exponent of 0.2, or a difference in surface curvature of 0.8 [Formula: see text] for surfaces with curvatures between 1 and 3 [Formula: see text]. In contrast, visual perception of similarity between color-coded images of the surface height was dominated by their topographic resemblance. We conclude that vibration cues from roughness at the length scale of the finger ridge distance distract the participants from including the topography into the judgement of similarity. The interaction between surface asperities and fingertip skin led to higher friction for higher micro-scale roughness. Individual friction data allowed us to construct a psychometric curve which relates similarity decisions to differences in friction. Participants noticed differences in the friction coefficient as small as 0.035 for samples with friction coefficients between 0.34 and 0.45.


Interprofessional health education teacher training at the University of Chile.
The first interprofessional course that included students in the 8 undergraduate health programs at the Faculty of Medicine of the University of Chile was implemented in 2015. For the 700 students, 35 teachers were trained as facilitators. The use of several strategies to train facilitators in interprofessional health education, such as working in small groups, role-playing, case analysis, personal development workshops with experts' participation, teamwork skills, feedback, videos, and reading articles, proved to be helpful. Facilitators highlighted the use of syllabi as a fundamental tool for teaching and coordination. This guide describes the experience of interprofessional health education teacher training from 2015 to 2019, highlighting the following lessons learned: the importance of support from university authorities, raising faculty awareness about interprofessional health education and collaborative practice, creating a teachers' coordination team including representatives from all health programs, and ongoing monitoring and feedback from participants.


Thiamine withdrawal can lead to diabetic ketoacidosis in thiamine responsive megaloblastic anemia: report of two siblings.
Thiamine responsive megaloblastic anemia syndrome (TRMA), an autosomal recessive disorder caused by the deficiency of thiamine transporter protein, is the association of diabetes mellitus, anemia and deafness. Pharmacological dose thiamine normalizes hematological abnormalities and their effects on the course of diabetes mellitus. We report on 8 years follow up of two siblings with TRMA. They presented in the prepubertal period with diabetic ketoacidosis due to lack of thiamine supplementation for 2 months. Their insulin requirements fell rapidly and disappeared with thiamine therapy. Hematological parameters normalized within 30 days. The diabetic picture is responsive to thiamine treatment in patients with TRMA. Insulin dependent diabetes may occur throughout the pubertal period. If thiamine supplementation is not sufficient, ketoacidosis may develop in patients during the prepubertal period.


Shear-induced alignment of block copolymer worms in mineral oil.
Poly(stearyl methacrylate)-poly(benzyl methacrylate) [PSMA-PBzMA] diblock copolymer worms were synthesized directly in mineral oil via reversible addition-fragmentation chain transfer (RAFT) dispersion polymerization at 90 °C. Free-standing gels were obtained from this polymerization-induced self-assembly (PISA) formulation when targeting PSMA13-PBzMA65 dispersions at 5% w/w to 20% w/w copolymer concentration. Gel permeation chromatography (GPC) studies indicated that almost identical copolymer chains were obtained in all cases, while transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS) studies confirmed that highly anisotropic worms were formed with mean cross-sectional diameters of 11.9-13.1 nm. These worms undergo a thermoreversible worm-to-sphere transition on heating up to 150 °C. Rheological studies were conducted to characterize the shear rate- and concentration-dependent behaviour caused by this change in copolymer morphology, where the initial shear-thinning worm gels form spheres (i.e. a Newtonian fluid) on heating up to 150 °C. Complementary shear-induced polarized light imaging (SIPLI) experiments confirmed the formation of aligned linear worms under applied shear between 80 °C and 110 °C, with high-viscosity dispersions of branched worms being obtained at 20-60 °C and low-viscosity spheres being produced at 150 °C. This study informs the use of such block copolymer worms as rheology modifiers for non-polar oils, which is of potential interest for the automotive industry.


Monoclonal antibodies as probes of high-density lipoprotein structure: identification and localization of a lipid-dependent epitope.
Eight stable murine monoclonal antibodies (mabs) were raised against human high-density lipoproteins (HDL). Three different antibody reactivities were demonstrated by immunoblotting. A group of five antibodies were specific for apolipoprotein A-I (apoA-I) and bound to similar or overlapping epitopes. The second type of reactivity, shown by mab-32, was specific for apoA-II. In the third group, two antibodies showed high reactivity with apoA-II and slight cross-reactivity with apoA-I. The properties of two antibodies, mab M-30 specific for apoA-I and mab M-32 specific for apoAII, were characterized in detail as probes of HDL structure. The association of 125I-labeled HDL or synthetic complexes of apoA-I and phosphatidylcholine with mab M-30 was lipid dependent. Mab M-32 binding to apoA-II was independent of lipid. The lipid-dependent epitope bound by mab M-30 has been localized to an 18 amino acid synthetic apoA-I peptide. Moreover, studies with HDL2, HDL3, and immunoadsorbed HDL subfractions indicate that binding of mab M-30 to HDL is influenced by some component within the microenvironment individual HDL particles. These lines of evidence suggest that the molar ratio of apoA-I to apoA-II is the critical determinant. Binding of mab M-32 to HDL increased the reactivity of HDL to mab M-30 in a dose-dependent manner, indicating an unusual form of cooperativity between two mabs that recognize different proteins in HDL. These monoclonal antibodies will be valuable in studies of the metabolic significance of protein-protein and lipid-protein interactions in HDL.


Biochemical properties and in planta effects of NopM, a rhizobial E3 ubiquitin ligase.
Nodulation outer protein M (NopM) is an IpaH family type three (T3) effector secreted by the nitrogen-fixing nodule bacterium Sinorhizobium sp. strain NGR234. Previous work indicated that NopM is an E3 ubiquitin ligase required for an optimal symbiosis between NGR234 and the host legume Lablab purpureus Here, we continued to analyze the function of NopM. Recombinant NopM was biochemically characterized using an in vitro ubiquitination system with Arabidopsis thaliana proteins. In this assay, NopM forms unanchored polyubiquitin chains and possesses auto-ubiquitination activity. In a NopM variant lacking any lysine residues, auto-ubiquitination was not completely abolished, indicating noncanonical auto-ubiquitination of the protein. In addition, we could show intermolecular ubiquitin transfer from NopM to C338A (enzymatically inactive NopM form) in vitro Bimolecular fluorescence complementation analysis provided clues about NopM-NopM interactions at plasma membranes in planta NopM, but not C338A, expressed in tobacco cells induced cell death, suggesting that E3 ubiquitin ligase activity of NopM induced effector-triggered immunity responses. Likewise, expression of NopM in Lotus japonicus caused reduced nodule formation, whereas expression of C338A showed no obvious effects on symbiosis. Further experiments indicated that serine residue 26 of NopM is phosphorylated in planta and that NopM can be phosphorylated in vitro by salicylic acid-induced protein kinase (NtSIPK), a mitogen-activated protein kinase (MAPK) of tobacco. Hence, NopM is a phosphorylated T3 effector that can interact with itself, with ubiquitin, and with MAPKs.


Cultured keratinocytes in in vitro dermatotoxicological investigation: a review.
The keratinocyte is responsible for the architecture of the epidermis, that portion of the skin that forms the environmental barrier necessary for survival. It also interacts with other cell types in the epidermis in response to various environmental influences. This cell type is used frequently for in vitro cutaneous toxicological investigations as an alternative to whole-animal studies. Several areas of cutaneous research using cultured keratinocytes are germane as regards the scope of this journal. The following areas of biomedical research were reviewed: (1) dermatotoxicology, including environmental chemicals, antiseptics, drugs, metals, and pesticides; (2) immunotoxicology, including inflammation and allergic dermatitis; (3) radiation, including ultraviolet and x-irradiation; and (4) the development of assays as alternatives to whole-animal testing. Due to the abundance of such investigations reported in the last 30 years, this review is limited mainly to reviewing reports published in this decade.


Blood lipids in children: age-related patterns and association with body-fat indices: Project HeartBeat!
Longitudinal data on the normal development of blood lipids and its relationships with body fatness in children and adolescents are limited. Objectives of the current analysis were to estimate trajectories related to age for four blood lipid components and to examine the impact of change in body fatness on blood lipid levels, comparing estimated effects among adiposity indices, in children and adolescents. Three cohorts, with a total of 678 children (49.1% female, 79.9% nonblack) initially aged 8, 11, and 14 years, were followed at 4-month intervals (1991-1995). Total cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride levels were determined in blood samples taken following fasting. Body fatness was measured by five adiposity indices-BMI; percent body fat (PBF); abdominal circumference; and the sums of six and of two skinfold thicknesses. Trajectories of change in blood lipid levels from ages 8 to 18 years were estimated by gender and race. The impact of change in body fatness on lipid levels was evaluated for each index, adjusting for gender, race, and age. All lipid components varied significantly with age. Total cholesterol decreased by approximately 19 mg/dL from ages 9 to 16 years in girls and more steeply from ages 10 to 17 years in boys. LDL-C decreased monotonically, more steeply in boys than in girls. It was higher among nonblacks than among blacks. HDL-C increased monotonically in girls, mainly from ages 14 to 18 years, but fluctuated sharply among boys. Levels of HDL-C were higher among blacks than among nonblacks. The levels of triglycerides increased from ages 8 to 12 years among girls and, almost linearly, from ages 8 to 18 years among boys. The levels of triglycerides were higher among nonblacks than among blacks. Increase in body fatness was significantly associated with increases in total cholesterol, LDL-C, and triglyceride levels. Significant interactions between the adiposity indices (except for BMI) and gender indicated smaller impacts of change in body fatness on total cholesterol and LDL-C in girls than in boys. The estimated impact on triglycerides was weaker among blacks than among nonblacks, except for PBF. Change in body fatness was negatively associated with HDL-C. The results remained essentially unchanged after adjustments for energy intake, physical activity, and sexual maturation. Patterns of change with age in blood lipid components vary significantly among gender and racial groups. Increase in body fatness among children is consistently associated with adverse change in blood lipids. Evaluation of blood lipid level should take into account variation by age, gender, and race. Intervention through body-fat control should help prevent adverse lipid levels in children and adolescents.


Histamine and Delirium: Current Opinion.
Delirium is a very common, but refractory clinical state, notably present in intensive care and in the growing aging community. It is characterized by fluctuating disturbances in a number of key behavioral features, namely cognition, mood, attention, arousal, and self-awareness. Histamine is arguably the most pleotropic neurotransmitter in the human brain, and this review provides a rationale, and proposes that this neuroactive amine plays a role in modulating the characteristic features of delirium. While centrally permeable H1 and H2 histamine receptor antagonists have pro-delirium potential, we propose that centrally permeable H3 histamine receptor antagonists may provide an exciting new strategy to combat delirium. The Histamine H4 receptor may also have an indirect inflammatory neuroglial role which requires further exploration.


When Is a Maze Procedure a Maze Procedure?
The initial surgical attempts to treat atrial fibrillation (AF) were isolation procedures designed to confine the arrhythmia to a specific area of the heart for relief of symptoms. The first surgical attempt to ablate AF was unsuccessful but was quickly followed by the Maze-I procedure on September 25, 1987. Because of several adverse sequelae of the Maze-I procedure, it was sequentially modified to the Maze-II and then Maze-III procedures. The Maze-IV procedure was introduced some 10 years later; these are the only 4 procedures that adhere to the concept of a maze pattern of lesions to ablate AF and leave both atria capable of being activated during normal sinus rhythm. The term maze procedure has become generic for virtually any operation designed to treat AF, but procedures that do not adhere to the concept of creating lesions of conduction block in the pattern of a maze are not maze procedures. These include, among others, the Wolf Mini-Maze, the Left-Sided Maze, and the 5-Box Maze, none of which are truly based on the maze-pattern concept. The cardinal feature of maze procedures that is necessary for both effectiveness and comparability to classical maze procedures includes lines of conduction block that preclude macro-reentry anywhere in either atrium while leaving both atria capable of activation by a sinus-generated impulse. Components essential to achieving this include appropriate lesions in both atria, the absence of gaps that allow electrical activity to bypass an intended line of block, and the absence of alternate pathways by which impulses can reach the intended maze exit.


Effects of glucagon and Ca2+ on the metabolism of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in isolated rat hepatocytes and plasma membranes.
Rat hepatocytes whose phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) had been labelled for 60 min with 32P were treated with glucagon for 10 min or phenylephrine for 2 min. Glucagon caused a 20% increase in PIP but no change in PIP2 whereas phenylephrine caused a similar increase in PIP but a 15% decrease in PIP2. Addition of both hormones together for 10 min produced a 40% increase in PIP. A crude liver mitochondrial fraction incubated with [32P]Pi and ADP incorporated label into PIP, PIP2 and phosphatidic acid. The PIP2 was shown to be in contaminating plasma membranes and PIP in both lysosomal and plasma-membrane contamination. A minor but definitely mitochondrial phospholipid, more polar than PIP2, was shown to be labelled with 32P both in vitro and in hepatocytes. The rate of 32P incorporation into PIP was faster in mitochondrial/plasma-membrane preparations from rats treated with glucagon or if 3 microM-Ca2+ and Ruthenium Red were present in the incubation buffer. Loss of 32P from membranes labelled in vitro was shown to be accompanied by formation of inositol 1,4,5-trisphosphate (IP3) and inositol 1,4-bisphosphate, and was faster in preparations from glucagon-treated rats or in the presence of 3 microM-Ca2+. It is concluded that glucagon stimulates both PIP2 phosphodiesterase and phosphatidylinositol kinase activities, as does the presence of 3 microM-Ca2+. The resulting formation of IP3 may be responsible for the observed release of intracellular Ca2+ stores. The roles of a guanine nucleotide regulatory protein and phosphorylation in mediating these effects are discussed.


XNF-ATc3 affects neural convergent extension.
Convergent extension is the primary driving force elongating the anteroposterior body axis. In Xenopus, convergent extension occurs in the dorsal mesoderm and posterior neural ectoderm, and is mediated by similar molecular pathways within these tissues. In this paper, we show that activation of NF-AT, a transcription factor known to modulate multiple signaling events, inhibits convergent extension in the dorsal mesoderm and in the posterior neural ectoderm. This is seen in whole embryos, mesodermal explants and posterior neural explants, solidly implicating a role of NF-AT in convergent extension. In the whole embryo, inhibition of NF-AT reveals a more selective function, affecting only convergent extension in the neural ectoderm. This specific activity was further teased apart using a variety of temporal and spatial approaches. Targeted injections of dominant-negative XNF-ATc3, or dosing over time with the calcineurin inhibitor cyclosporin in neural tube explants or in whole embryos, shows that inhibition of NF-AT signaling blocks neural convergent extension. Consistent with a function in neural convergent extension, we show that XNF-ATc3 is expressed and transcriptionally active within the neural tube. This work identifies XNF-ATc3 as a regulator of neural convergent extension in Xenopus and adds to a short list of molecules involved in this process.


Ex vivo Hematopoietic Stem Cell Expansion Technologies: Recent Progress, Applications, and Open Questions.
Hematopoietic stem cells (HSCs) are a rare but potent cell type that support life-long hematopoiesis and will stably regenerate the entire blood and immune system following transplantation. HSC transplantation represents a mainstay treatment for various diseases of the blood and immune systems. The ex vivo expansion and manipulation of HSCs therefore represents an important approach to ask biological questions in experimental hematology and to help improve clinical HSC transplantation therapies. However, it has remained challenging to expand transplantable HSCs ex vivo. This review summarizes recent progress in ex vivo HSC expansion technologies and their applications to biological and clinical problems, and discusses current questions in the field.


Surface-confined photopolymerization of pH-responsive acrylamide/acrylate brushes on polymer thin films.
Dynamic acrylamide/acrylate polymeric brushes were synthesized at gold-plated quartz crystal surfaces. The crystals were initially coated with polystyrene-type thin films, derivatized with photolabile iniferter groups, and subsequently subjected to photoinitiated polymerization in acrylamide/acrylate monomer feeds. This surface-confined polymerization method enabled direct photocontrol over the polymerization, as followed by increased frequency responses of the crystal oscillations in a quartz crystal microbalance (QCM). The produced polymer layers were also found to be highly sensitive to external acid/base stimuli. Large oscillation frequency shifts were detected when the brushes were exposed to buffer solutions of different pH. The dynamic behavior of the resulting polymeric brushes was evaluated, and the extent of expansion and contraction of the films was monitored by the QCM setup in situ in real time. The resulting responses were rapid, and the effects were fully reversible. Low pH resulted in full contractions of the films, whereas higher pH yielded maximal expansion in order to minimize repulsion around the charged acrylate centers. The surfaces also proved to be very robust because the responsiveness was reproducible over many cycles of repeated expansion and contraction. Using ellipsometry, copolymer layers were estimated to be approximately 220 nm in a collapsed state and approximately 340 nm in the expanded state, effectively increasing the thickness of the film by 55%.


Surveillance of Echinococcus granulosus in dogs with arecoline hydrobromide.
Arecoline hydrobromide has been used for almost half a century for the treatment of Echinococcus granulosus in dogs. Trials in New Zealand showed that it had real limitations for this purpose. Its main value lies in its use as a diagnostic agent for detecting infections in dogs on a group basis. The data so obtained can be used in educating dog owners as well as for providing base-line data and an index of progress in a continuing control programme. The drug's limitations for treatment and value as a diagnostic agent in a field trial are assessed.


Neutrophil-derived TNF-related apoptosis-inducing ligand (TRAIL): a novel mechanism of antitumor effect by neutrophils.
To detect the novel genes expressed uniquely in neutrophils and elucidate their function, the gene expression pattern was compared by using cDNA microarray containing 240 cytokine genes between the neutrophils and peripheral blood mononuclear cells (PBMCs) obtained from healthy human donors. Twenty-six genes, including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), were expressed in neutrophils at a level >10 times higher than that seen in phytohemagglutinin-stimulated PBMCs. The amounts of mRNA and protein of TRAIL were quantified by real-time reverse transcription-PCR and ELISA, respectively. TRAIL was expressed in resting neutrophils at the mRNA and protein levels, and its expression was enhanced after stimulation with IFN-gamma. Neutrophils expressed TRAIL on the cell surface and released it into the culture media. The cytotoxicity of neutrophil-derived TRAIL against Jurkat cells was determined by flow cytometry using FITC-conjugated annexin V. When Jurkat cells were cultured with neutrophils in the presence of IFN-gamma, the number of Jurkat cells undergoing apoptosis increased, and such increase depended on the effector:target ratio. This cytotoxicity was suppressed partially by adding anti-TRAIL antibody to the media. Neutrophils may exert their own antitumor effect by TRAIL. A microarray analysis was found to be a useful tool for detecting novel genes that are suggested to play unknown roles in the neutrophil function.


An evolutionary approach for image segmentation.
The paper explores the use of evolutionary techniques in dealing with the image segmentation problem. An image is modeled as a weighted undirected graph, where nodes correspond to pixels, and edges connect similar pixels. A genetic algorithm that uses a fitness function based on an extension of the normalized cut criterion is proposed. The algorithm employs the locus-based representation of individuals, which allows for the partitioning of images without setting the number of segments beforehand. A new concept of nearest neighbor that takes into account not only the spatial location of a pixel, but also the affinity with the other pixels contained in the neighborhood, is also defined. Experimental results show that our approach is able to segment images in a number of regions that conform well to human visual perception. The visual perceptiveness is substantiated by objective evaluation methods based on uniformity of pixels inside a region, and comparison with ground-truth segmentations available for part of the used test images.


[Mating types in the ciliate Dileptus anser. Inheritance and genetic determination].
Hybridological analysis of mating types (MTs) has been first made for the lower ciliate Dileptus anser. Clones of an initially unknown genotype belonging to three MTs (MT I, MT II and MT III), characteristic of D. anser, were isolated from natural reservoirs and further used for crosses. In one group crosses, synclonal inheritance and typical Mendelian behaviour of the character were observed over sexual generations of ciliates. The results suggest that MTs in D. anser may be directly controlled by a single mat locus with three alleles showing peck-order dominance (mat1 > mat2 > mat3). In other words, cells with mat1/mat1, mat1/mat2 and mat1/mat3 genotypes belong to MT I, those with mat2/mat2 and mat2/mat3, and the mat3/mat3 belong to MT II and MT III, respectively. Sexually mature exconjugant clones stably retain their MTs corresponding to their genotypes on vegetative reproduction. The progeny of other group crosses showed various deviations from typical Mendelian behaviour of the character. In some cases, standard Mendelian ratios were more or less violated. Most typical was instability of differentiation for MT in maturing exconjugant clones. Shortly after their maturation, the majority of clones change their MT, rather frequently more than once, although the finally established MT is stably inherited afterwards, during vegetative reproduction. When unstable, exconjugant clones can successively express two or even three MTs characteristic of this species, including MTs that should not have been expected on the basis of parental genotypes available in a given cross. It looks likely that the mat locus in D. anser is complex and multipotential; it is inherited as a whole providing for expression of any MT characteristic of the species (in this respect bearing similarity with Tetrahymena thermophila). Other mechanisms, epigenetic in particular (Nanney, 1958), determine the final expression of one of the three MT potentialities by a given exconjugant clone. Stable, persistent functioning of these mechanisms ensures a stable differentiation for MT and Mendelian behaviour of the character in sexual generations and in crosses. Any disturbances in differentiation control may trigger MT instability in maturing exconjugant clones and violation of regular Mendelian behaviour.


Echocardiographic features of total anomalous pulmonary venous connection.
Seven patients, four months to ten years of age, with total anomalous pulmonary venous connection (TAPVC) were studied by echocardiography. The diagnosis was confirmed by cardiac catheterization, angiography, and surgery in each. The connection was to the left vertical vein in four patients, to the coronary sinus in one, to the inferior vena cava in one, and directly to the right atrium in one. All patients exhibited the echocardiographic criteria of right ventricular diastolic volume overload (RVDVO). In addition, an echo-free space was identified dorsal to the posterior wall of the left atrium. Indocyanine green dye studies done in one of the patients provided evidence that this echo-free space represents the common pulmonary venous chamber. The combination of echocardiographic evidence of RVDVO and demonstration of an echo-free space dorsal to the left atrium is strongly suggestive of TAPVC and should allow an early diagnosis in infants with this anomaly.


The quality of sleep and daytime sleepiness and their association with academic achievement of medical students in the eastern province of Saudi Arabia.
Sleep is often challenging for medical students because of the considerable amount of learning they have to do. The aim of this study was to identify the patterns of sleep quality and daytime sleepiness among medical students in Saudi Arabia and to identify the possible factors associated with poor quality of sleep and daytime sleepiness. This cross-sectional questionnaire-based study was conducted at College of Medicine, Imam Abdulrahman Bin Faisal University, Saudi Arabia. The questionnaire included the prevalidated Pittsburgh Sleep Quality Index (PSQI) to measure sleep quality and the Epworth Sleepiness Scale (ESS) to measure daytime sleepiness. Covariates included sociodemographic characteristics and academic performance. Bivariate and multivariate logistic regression analyses were used to obtain adjusted and unadjusted odds ratios (ORs). The sampled population comprised 36.7% males and 63.30% females. The mean global PSQI score was (standard deviation (SD)=4.66), while the mean global ESS score was 8.03 (SD=4.66). Further analysis showed that 80.60% had poor sleep quality and 37.80% of the students suffered from excessive daytime sleepiness. Multivariate regression showed that poor sleep quality was significantly associated with poor academic achievement (adjusted OR = 3.33, 95% confidence interval [CI] = 1.28-8.63). Furthermore, excessive sleepiness significantly increased the odds of poor academic performance (adjusted OR = 4.58, 95% CI = 1.09-20.81). The results on daytime sleepiness were found to be within the ranges reported worldwide; poor sleep quality was found to be among the highest ever reported. Both sleep quality and daytime sleepiness significantly affect academic achievement.


Salvianolic acid B inhibits oxidative stress in glomerular mesangial cells alleviating diabetic nephropathy by regulating SIRT3/FOXO1 signaling.
Oxidative stress is pivotal in advancing diabetic nephropathy (DN). Salvianolic acid B (SAB), derived from Radix Salviae miltiorrhizae, exhibits renoprotective effects. However, the mechanisms underlying its action in DN are not fully elucidated. This study explores SAB's protective effect on DN, focusing on its antioxidative properties in glomerular mesangial cells. The renoprotective effects of various SAB dosages on DN rats were assessed by evaluating kidney tissue pathological alterations through hematoxylin-eosin (H&E), periodic acid-Schiff (PAS), Masson, TUNEL staining and kidney function through biochemical detection. CCK8 and lactate dehydrogenase (LDH) cytotoxicity assays were utilized to evaluate the viability of high glucose (HG)-induced HBZY-1 cells treated with various SAB dosages. Oxidative stress and inflammation levels were measured using enzyme-linked immunosorbent assay (ELISA) kits. The Sirtuin 3 (SIRT3)/Forkhead box transcription factor O1 (FOXO1) pathway was examined through Western blot and immunohistochemistry. SAB mitigated kidney histopathological alterations and function and cell apoptosis in DN rats at various dosages. It enhanced the activity of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) while decreasing reactive oxygen species (ROS) and malondialdehyde (MDA) levels both in vivo and in vitro. SAB also suppressed the levels of pro-inflammatory cytokines (IL-1β, IL-6, MCP-1, and TNF-α) and the expression of collagen IV and fibronectin in HG-induced HBZY-1 cells. Furthermore, SAB activated the SIRT3/FOXO1 signaling pathway. Our findings suggest that SAB may alleviate oxidative stress in DN both in vivo and in vitro, potentially through the activation of the SIRT3/FOXO1-mediated signaling pathway. This study provides initial insights into the possible antioxidative and renoprotective effects of SAB, indicating its potential utility as a therapeutic agent for DN.