Adaptive estimation and control method for unstable periodic dynamics in spike trains.
Dynamical control of excitable biological systems is often complicated by the difficult and unreliable task of precontrol identification of unstable periodic orbits (UPO's). Here we show that, for both chaotic and nonchaotic systems, UPO's can be located, and their dynamics characterized, during control. Tracking of system nonstationarities emerges naturally from this approach. Such a method is potentially valuable for the control of spike trains of excitable biological systems, for which precontrol UPO identification is often impractical, and nonstationarities (natural or stimulation induced) are common.


Influence of Axial Depth of Cut and Tool Position on Surface Quality and Chatter Appearance in Locally Supported Thin Floor Milling.
Thin floor machining is a challenging and demanding issue, due to vibrations that create poor surface quality. Several technologies have been developed to overcome this problem. Ad hoc fixtures for a given part geometry lead to meeting quality tolerances, but since they lack flexibility, they are expensive and not suitable for low manufacturing batches. On the contrary, flexible fixtures consisting of vacuum cups adaptable to a diversity of part geometries may not totally avoid vibrations, which greatly limits its use. The present study analyses the feasibility of thin floor milling in terms of vibration and roughness, in the cases where milling is conducted without back support, a usual situation when flexible fixtures are employed, so as to define the conditions for a stable milling in them and thus avoid the use of ad hoc fixtures. For that purpose, the change of modal parameters due to material removal and its influence on chatter appearance have been studied, by means of stability lobe diagrams and Fourier Transform analysis. Additionally, the relationship between surface roughness and chatter frequency, tooth passing frequency, and spindle frequency have been studied. Ploughing effect has also been observed during milling, and the factors that lead to the appearance of this undesirable effect have been analyzed, in order to avoid it. It has been proven that finish milling of thin floors without support in the axial direction of the mill can meet aeronautic tolerances and requirements, providing that proper cutting conditions and machining zones are selected.


Base-Promoted Synthesis of S-Arylisothiazolones via Intramolecular Dehydrative Cyclization of α-Keto-N-acylsulfoximines.
A base (Et3N)-promoted synthesis of 1,4-diarylisothiazolones from α-keto-N-acylsulfoximines has been achieved. The reaction proceeds via α-hydrogen abstraction from sulfoximine, followed by an intramolecular nucleophilic attack at the keto carbonyl to form a tert-hydroxy isothiazolone intermediate. The 1,4-substituted isothiazolone is obtained after dehydration via an E1cB path. This one-pot synthesis of isothiazolinones has a broad substrate scope, has a high atom economy, and provides products with good yields. The ΔELUMO-HOMO is calculated using Gaussian 16 at the B3LYP/6-31G(d,p) level of theory.


NKp46 and NKG2D recognition of infected dendritic cells is necessary for NK cell activation in the human response to influenza infection.
At an early phase of viral infection, contact and cooperation between dendritic cells (DCs) and NK cells activates innate immunity, and also influences recruitment, when needed, of adaptive immunity. Influenza, an adaptable fast-evolving virus, annually causes acute, widespread infections that challenge the innate and adaptive immunity of humanity. In this study, we dissect and define the molecular mechanisms by which influenza-infected, human DCs activate resting, autologous NK cells. Three events in NK cell activation showed different requirements for soluble mediators made by infected DCs and for signals arising from contact with infected DCs. IFN-alpha was mainly responsible for enhanced NK cytolysis and also important for CD69 up-regulation, whereas IL-12 was necessary for enhancing IFN-gamma production. Increased CD69 expression and IFN-gamma production, but not increased cytolysis, required recognition of influenza-infected DCs by two NK cell receptors: NKG2D and NKp46. Abs specific for these receptors or their known ligands (UL16-binding proteins 1-3 class I-like molecules for NKG2D and influenza hemagglutinin for NKp46) inhibited CD69 expression and IFN-gamma production. Activation of NK cells by influenza-infected DCs and polyinosinic:polycytidylic acid (poly(I:C))-treated DCs was distinguished. Poly(I:C)-treated DCs did not express the UL16-binding protein 3 ligand for NKG2D, and in the absence of the influenza hemagglutinin there was no involvement of NKp46.


A novel nonhuman primate model for influenza transmission.
Studies of influenza transmission are necessary to predict the pandemic potential of emerging influenza viruses. Currently, both ferrets and guinea pigs are used in such studies, but these species are distantly related to humans. Nonhuman primates (NHP) share a close phylogenetic relationship with humans and may provide an enhanced means to model the virological and immunological events in influenza virus transmission. Here, for the first time, it was demonstrated that a human influenza virus isolate can productively infect and be transmitted between common marmosets (Callithrix jacchus), a New World monkey species. We inoculated four marmosets with the 2009 pandemic virus A/California/07/2009 (H1N1pdm) and housed each together with a naïve cage mate. We collected bronchoalveolar lavage and nasal wash samples from all animals at regular intervals for three weeks post-inoculation to track virus replication and sequence evolution. The unadapted 2009 H1N1pdm virus replicated to high titers in all four index animals by 1 day post-infection. Infected animals seroconverted and presented human-like symptoms including sneezing, nasal discharge, labored breathing, and lung damage. Transmission occurred in one cohabitating pair. Deep sequencing detected relatively few genetic changes in H1N1pdm viruses replicating in any infected animal. Together our data suggest that human H1N1pdm viruses require little adaptation to replicate and cause disease in marmosets, and that these viruses can be transmitted between animals. Marmosets may therefore be a viable model for studying influenza virus transmission.


Cbln1 is a ligand for an orphan glutamate receptor delta2, a bidirectional synapse organizer.
Cbln1, secreted from cerebellar granule cells, and the orphan glutamate receptor delta2 (GluD2), expressed by Purkinje cells, are essential for synapse integrity between these neurons in adult mice. Nevertheless, no endogenous binding partners for these molecules have been identified. We found that Cbln1 binds directly to the N-terminal domain of GluD2. GluD2 expression by postsynaptic cells, combined with exogenously applied Cbln1, was necessary and sufficient to induce new synapses in vitro and in the adult cerebellum in vivo. Further, beads coated with recombinant Cbln1 directly induced presynaptic differentiation and indirectly caused clustering of postsynaptic molecules via GluD2. These results indicate that the Cbln1-GluD2 complex is a unique synapse organizer that acts bidirectionally on both pre- and postsynaptic components.


Differential expression of the intermediate filament protein nestin during renal development and its localization in adult podocytes.
Nestin, an intermediate filament protein, is widely used as stem cell marker. Nestin has been shown to interact with other cytoskeleton proteins, suggesting a role in regulating cellular cytoskeletal structure. These studies examined renal nestin localization and developmental expression in mice. In developing kidney, anti-nestin antibody revealed strong immunoreactivity in vascular cleft of the S-shaped body and vascular tuft of capillary loop-stage glomerulus. The nestin-positive structures also were labeled by endothelial cell markers FLK1 and CD31 in immature glomeruli. Nestin was not detected in epithelial cells of immature glomeruli. In contrast, in mature glomerular, nestin immunoreactivity was observed only outside laminin-positive glomerular basement membrane, and co-localized with nephrin, consistent with podocyte nestin expression. In adult kidney, podocytes were the only cells that exhibited persistent nestin expression. Nestin was not detected in ureteric bud and its derivatives throughout renal development. Cell lineage studies, using a nestin promoter-driven Cre mouse and a ROSA26 reporter mouse, showed a strong beta-galactosidase activity in intermediate mesoderm in an embryonic day 10 embryo and all of the structures except those that were derived from ureteric bud in embryonic kidney through adult kidney. These studies show that nestin is expressed in progenitors of glomerular endothelial cells and renal progenitors that are derived from metanephric mesenchyme. In the adult kidney, nestin expression is restricted to differentiated podocytes, suggesting that nestin could play an important role in maintaining the structural integrity of the podocytes.


Experimental pain in the groin may refer into the lower abdomen: Implications to clinical assessments.
To investigate the effects of experimental adductor pain on the pain referral pattern, mechanical sensitivity and muscle activity during common clinical tests. Repeated-measures design. In two separate sessions, 15 healthy males received a hypertonic (painful) and isotonic (control) saline injection to either the adductor longus (AL) tendon to produce experimental groin pain or into the rectus femoris (RF) tendon as a painful control. Pain intensity was recorded on a visual analogue scale (VAS) with pain distribution indicated on body maps. Pressure pain thresholds (PPT) were assessed bilaterally in the groin area. Electromyography (EMG) of relevant muscles was recorded during six provocation tests. PPT and EMG assessment were measured before, during and after experimental pain. Hypertonic saline induced higher VAS scores than isotonic saline (p<0.001), and a local pain distribution in 80% of participants. A proximal pain referral to the lower abdominal region in 33% (AL) and 7% (RF) of participants. Experimental pain (AL and RF) did not significantly alter PPT values or the EMG amplitude in groin or trunk muscles during provocation tests when forces were matched with baseline. This study demonstrates that AL tendon pain was distributed locally in the majority of participants but may refer to the lower abdomen. Experimental adductor pain did not significantly alter the mechanical sensitivity or muscle activity patterns.


Influence of conductance changes on patch clamp capacitance measurements using a lock-in amplifier and limitations of the phase tracking technique.
We characterized the influence of conductance changes on whole-cell patch clamp capacitance measurements with a lock-in amplifier and the limitations of the phase-tracking method by numerical computer simulations, error formulas, and experimental tests. At correct phase setting, the artifacts in the capacitance measurement due to activation of linear conductances are small. The cross talk into the capacitance trace is well approximately by the second-order term in the Taylor expansion of the admittance. In the case of nonlinear current-voltage relationships, the measured conductance corresponds to the slope conductance in the range of the sine wave amplitude, and the cross talk into the capacitance trace corresponds to the second-order effect of the slope conductance. The finite gating kinetics of voltage-dependent channels generate phase-shifted currents. These lead to major artifacts in the capacitance measurements when the angular frequency of the sine wave is close to the kinetic rate constant of the channel. However, when the channel kinetics are sufficiently slow, or sufficiently fast, the cross talk is still close to the second-order effect of the measured conductance. The effects of activation of voltage-dependent currents on the capacitance measurements may be estimated, provided a detailed characterization of the kinetics and voltage dependence is available. A phase error of the lock-in amplifier of a few degrees leads to significant projections. The phase-tracking method can be used to keep the phase aligned only during periods of low membrane conductance. However, nonideal properties of the equivalent circuit, in particular the fast capacitance between the pipette and the bath solutions, may lead to large phase errors when the phase-tracking method is used, depending on the electrical properties of the cell. In this article we provide practical values, setting the range where possible artifacts are below defined limits. For proper evaluation of capacitance measurements, the capacitance and conductance traces should always be displayed together.


Adrenocortical carcinoma: Diagnosis, prognostic classification and treatment of localized and advanced disease.
Adrenocortical carcinoma (ACC) is a rare cancer with an estimated incidence of 0.7 to 2.0 cases per 1 million population per year in the United States. It is an aggressive cancer originating in the cortex of the adrenal gland with a poor prognosis. The 5-year survival rate is less than 15% among patients with metastatic disease. In this article, we review the epidemiology and pathogenesis of ACC, the diagnostic procedures, the prognostic classification of ACC, and the treatment options from localized and resectable forms to advanced disease detailing recent therapeutic developments such as immunotherapy and molecularly targeted therapy.


Relaparoscopy in minor bile leakage after laparoscopic cholecystectomy: an alternative approach?
Bile leakage is one of the most important complications of laparoscopic cholecystectomy, and it has been recognized as a major clinical challenge during the last 2 decades. Although endoscopic and percutaneous interventions are widely accepted in the management of bile leakage, relaparoscopy permits the bile drainage adequately and gives direct control of bile leakage site in selected patients. Data for patients with minor bile leakage after laparoscopic cholecystectomy were collected from January 2001 to January 2010. Patients were categorized in 2 groups, nonoperative and relaparoscopy. Clinical presentation, kind of management, and outcomes were evaluated in 2 groups. After a total of 2652 laparoscopic cholecystectomies, postoperative minor bile leakage occurred in 17 (0.64%) patients. Four patients with minimal leakage were managed by percutaneous drainage alone. Endoscopic retrograde cholangiopancreatography was applied to 4 patients with jaundice, high output bile fistula, and a patient with retained common bile duct stone. Bile leakage was controlled in 3 of the 4 patients. There were 9 patients in the relaparoscopic group to which 1 patient was added after unsuccessful endoscopic intervention. The source of bile leakage in the relaparoscopic cases was defined as 50% from cystic duct stump and 50% from Luschka or accessory ducts. The success rate of bile leakage control after relaparoscopy was 90%. The mean of hospital stay after relaparoscopy was 3 days (range, 2-10 d) and after endoscopic retrograde cholangiopancreatography intervention or percutaneous drainage was 10 days (range, 3-28 d). Relaparoscopy is an effective procedure in the management of minor bile leakage after laparoscopic cholecystectomy and can be an alternative approach in selected situations.


[Pneumatosis cystoides coli--a rare differential diagnosis in colonoscopy].
Pneumatosis cystoides intestinalis (PCI), also called emphysema intestinalis or colitis cystica is a rare entity of unknown etiology. Characteristic signs can be found in endoscopy. We report the case of a 63 years old man in whom the misinterpretation of the colonoscopic signs resulted in resection of the ascending colon. After surgical therapy the patient is up to now asymptomatic with no signs of recurrence. Treatment of PCI is necessary only in symptomatic cases. The first choice of treatment is oxygen therapy in combination with metronidazole for six weeks. High recurrence rates are found with either oxygen treatment or antibiotic application alone. Surgical therapeutic intervention is recommended only in cases with serious complications. Limited resection of the affected part of the bowel should be carried out.


Species delimitation using Bayes factors: simulations and application to the Sceloporus scalaris species group (Squamata: Phrynosomatidae).
Current molecular methods of species delimitation are limited by the types of species delimitation models and scenarios that can be tested. Bayes factors allow for more flexibility in testing non-nested species delimitation models and hypotheses of individual assignment to alternative lineages. Here, we examined the efficacy of Bayes factors in delimiting species through simulations and empirical data from the Sceloporus scalaris species group. Marginal-likelihood scores of competing species delimitation models, from which Bayes factor values were compared, were estimated with four different methods: harmonic mean estimation (HME), smoothed harmonic mean estimation (sHME), path-sampling/thermodynamic integration (PS), and stepping-stone (SS) analysis. We also performed model selection using a posterior simulation-based analog of the Akaike information criterion through Markov chain Monte Carlo analysis (AICM). Bayes factor species delimitation results from the empirical data were then compared with results from the reversible-jump MCMC (rjMCMC) coalescent-based species delimitation method Bayesian Phylogenetics and Phylogeography (BP&P). Simulation results show that HME and sHME perform poorly compared with PS and SS marginal-likelihood estimators when identifying the true species delimitation model. Furthermore, Bayes factor delimitation (BFD) of species showed improved performance when species limits are tested by reassigning individuals between species, as opposed to either lumping or splitting lineages. In the empirical data, BFD through PS and SS analyses, as well as the rjMCMC method, each provide support for the recognition of all scalaris group taxa as independent evolutionary lineages. Bayes factor species delimitation and BP&P also support the recognition of three previously undescribed lineages. In both simulated and empirical data sets, harmonic and smoothed harmonic mean marginal-likelihood estimators provided much higher marginal-likelihood estimates than PS and SS estimators. The AICM displayed poor repeatability in both simulated and empirical data sets, and produced inconsistent model rankings across replicate runs with the empirical data. Our results suggest that species delimitation through the use of Bayes factors with marginal-likelihood estimates via PS or SS analyses provide a useful and complementary alternative to existing species delimitation methods.


Broadband EMFi-based transducers for ultrasonic air applications.
In this work, we explore the possibilities of electromechanical film (EMFi) as a new material for developing broadband transducers for ultrasonic air applications. The advantages of the EMFi film are its wide usable frequency range and easiness to use, making it highly suitable for self made, customizable ultrasonic sensors. This paper presents theoretical and experimental information focused on the needs of the sensor's end user, namely, frequency response, actual dynamic mass and Young's modulus, bandwidth, sensitivity, electromechanical dynamical model, acoustic response, and directivity. It is found empirically that the behavior of the film as an almost ideal piston-like acoustic source permits accurate prediction of the characteristics of transducers built on a developable surface. The results obtained represent the first step to more complex geometries, and, ultimately, to completely customizable field ultrasonic transducers.


The history of surgery according to Owen Wangensteen.
Owen Wangensteen (1898-1981), the great Minnesota surgical master, made a firm and intense dedication to the history of surgery from the early stages of his exemplary surgical career. While in Germany and Switzerland, he learned from distinguished European professors and clearly realized the importance of history on the understanding and appreciation of important surgical problems. In 1928, on returning to Minneapolis, he began his quest to introduce history into many of his clinical and research laboratory works. In 1930, he attained the surgery chairmanship at the University of Minnesota, and there he excelled until his retirement from surgical practice in 1967. Throughout his career and until he died in 1981, the history of surgery was his continuous and persistent companion. In fact, this discipline, the history of surgery, was indeed part of the family, since his wife Sally Davidson Wangensteen was a dedicated contributor to their common papers and the scholarly jewel of their work, the monumental treatise, The Rise of Surgery: From Empiric Craft to Scientific Discipline.


Imaging Diagnosis of Primary Solitary Bone Neoplasts and Its Comparison with Tumor-like Lesions.
To explore the imaging diagnostic value of primary solitary bone tumor and tumor-like lesion of iliac crest. A total of 156 patients with primary solitary bone tumors and tumor-like lesions of the iliac bone treated in our hospital were selected, and the patients were diagnosed by X-ray, CT, and MRI. Sexual analysis of single diagnostic and combined diagnostic value was carried out. Round high-density shadow, soft tissue mass shadow, soft tissue mass, right intestinal tube, and bladder obvious pressure were observed. The detection rates of giant cell tumor of bone, myeloma, osteochondroma, chondroma, eosinophilic granuloma, osteosarcoma, fibrous dysplasia, and Hodgkin lymphoma were 34.6%, 12.8%, 11.5%, 10.3%, 7.7%, 6.4%, 3.8%, and 2.6%, and the differences were statistically significant (P < 005); X-ray, CT, MR single diagnostic comparison, three methods joint diagnostic missed diagnosis rate and misdiagnosis rate, higher detection rate (P < 0.05); combined with X-ray, CT, MR single diagnosis, three methods joint diagnosis sensitivity, specificity, accuracy, statistical significance (P < 0.05); comparison with X-ray, CT, MR single diagnosis, three methods jointly diagnosed positive predictive value, negative predictive value higher, difference statistics significance (P < 0.05); there is a significant difference in the near-end, backbone, and distal detection rate of different bone tumors and tumor lesions, including the humerus and tibia. There is a statistical significance of the detection rate, and the difference isP < 0.05. X-ray plays an important role in the diagnosis of primary solitary bone tumor and tumor-like lesion of iliac crest and is the first choice in clinical diagnosis. In the diagnosis of tumor disease, range, and soft tissue mass, MRI and CT diagnostic value can provide effective theoretical basis for patient clinical treatment. Therefore, the appropriate diagnostic method should be selected according to the specific situation of the patient, so that the efficiency of the clinical feature is improved.


Long term life dissatisfaction and subsequent major depressive disorder and poor mental health.
Poor mental health, especially due to depression, is one of the main public health problems. Early indicators of poor mental health in general population are needed. This study examined the relationship between long-term life dissatisfaction and subsequent mental health, including major depressive disorder. Health questionnaires were sent to a randomly selected population-based sample in 1998 and repeated in 1999 and 2001. In 2005, a clinically studied sub-sample (n = 330) was composed of subjects with (n = 161) or without (n = 169) repeatedly reported adverse mental symptoms at all three previous data collection times. Clinical symptom assessments were performed with several psychometric scales: life satisfaction (LS), depression (HDRS, BDI), hopelessness (HS), mental distress (GHQ), dissociative experiences (DES), and alexithymia (TAS). The long-term life dissatisfaction burden was calculated by summing these life satisfaction scores in 1998, 1999, 2001 and dividing the sum into tertiles. Psychiatric diagnoses were confirmed by SCID-I for DSM-IV in 2005. Logistic regression analyses were performed to assess the studied relationship. The previous life dissatisfaction burden associated with adverse socio-demographic, life style and clinical factors. In adjusted logistic regression analyses, it was independently and strongly associated with subsequent major depressive disorder in 2005, even when the concurrent LS score in 2005 was included in the model. Excluding those with reported major depressive disorder in 1999 did not alter this finding. MDD in 1999 was based on self-reports and not on structured interview and LS data in 2001-2005 was not available. The life satisfaction burden is significantly related to major depressive disorder and poor mental health, both in cross-sectional and longitudinal settings.


Transfer of glycosylphosphatidylinositol-anchored proteins to deficient cells after erythrocyte transfusion in paroxysmal nocturnal hemoglobinuria.
In paroxysmal nocturnal hemoglobinuria (PNH), an acquired mutation of the PIGA gene results in the absence of glycosylphosphatidylinositol (GPI)-anchored cell surface membrane proteins in affected hematopoietic cells. Absence of GPI-anchored proteins on erythrocytes is responsible for their increased sensitivity to complement-mediated lysis, resulting in hemolytic anemia. Cell-to-cell transfer of CD55 and CD59, 2 GPI-anchored proteins, by red cell microvesicles has been demonstrated in vitro, with retention of their function. Because red cell units stored for transfusion contain many erythrocyte microvesicles, transfused blood could potentially serve as a source of CD55 and CD59. We examined whether GPI-anchored proteins could be transferred in vivo to deficient cells following transfusions given to 6 patients with PNH. All patients were group A(1) blood type. Each was given transfusions of 3 U of compatible, washed group O blood. Patient group A(1) cells were distinguished from the transfused group O cells by flow cytometry and staining with a labeled lectin, Dolichos biflorus, which specifically binds to group A(1) erythrocytes. Increased surface CD59 was measured on recipient red cells and granulocytes 1, 3, and 7 days following transfusion in all 6 patients. Our data suggest a potential therapeutic role for GPI-anchored protein transfer for severe PNH.


[Effect of T-activin on peripheral immunity organs in intact and thymectomized mice].
Morphometry of the spleen, axillary lymph nodes and cytological assay of the bone marrow and peripheral blood were performed in (CBA X C57BL)F1 mice 1, 5, 10 and 15 days after subcutaneous injection of 0.5 microgram T-activin to intact and thymectomized (when adult) mice 2 months after operation. It was demonstrated that in intact animals, injection of T-activin stimulated the whole system of immunogenesis. The time course of plasmatization and the response of the germinative centers differing from that seen during antigen administration suggests that T-activin is not immunogenous, acting as a stimulant of the previous immune responses. The permanent amount of the degenerating cells attests to the lack of the toxic drug effect.


Effect of Pistacia atlantica kurdica gum on diabetic gastroparesis symptoms: a randomized, triple-blind placebo-controlled clinical trial.
Gastroparesis is a recognized problem of type 1 and 2 diabetes mellitus that is defined by delayed gastric emptying. Due to insufficient satisfaction of current treatments, the use of complementary and alternative medicine in this field seems essential. According to Persian medicine, Pistacia atlantica gum has proper effect on gastrointestinal disorder specially stomach diseases. The aim of this study was to assess the effect of Pistacia atlantica gum on diabetic gastroparesis symptoms. A triple-blind randomized placebo-controlled trial on subjects with diabetic gastroparesis was conducted at the clinic of Imam Reza Hospital in Mashhad city in Iran, from November 2016 to May 2017. Fifty-four patients (16.7% males and 83.3% females) enrolled in the 8-week study and were randomly divided into two groups of intervention Pistacia atlantica and placebo groups. Both of the groups had received current dietary advice. Severity of gastroparesis symptoms was assessed with a Gastroparesis cardinal symptom index (GCSI) questionnaire. SPSS software version 16 was used to analyze the data and Generalized Estimating Equation (GEE) model, Independent-samples t-test, Paired-samples t-test, Mann-Whitney U test, and Wilcoxon signed-rank test were used to assess variables. All gastroparesis symptoms of patients (14.6% diabetes type 1 and 85.4% diabetes type 2) including nausea/vomiting, postprandial fullness/early satiety, and bloating were significantly declined in each group during study (p<0.0001). Pistacia atlantica gum significantly reduced severity of gastroparesis symptoms including Postprandial fullness/early satiety (p=0.04), Bloating (p<0.0001), and GCSI scores (p=0.001) versus placebo. Systolic blood pressure (p=0.006), BMI (p=0.018), and HbA1c (p=0.038) significantly decreased in the Pistacia atlantica group after the intervention. This study showed that daily consumption of Pistacia atlantica gum for one month improves diabetic gastroparesis symptoms. The trial was registered at the Iranian Registry of Clinical Trials (http://www.irct.ir) with the code of IRCT2016110630739N1. This study was supported by funding (grant no.: 950153) from the Mashhad University of Medical Sciences Research Council, Mashhad, Iran.


Differential cytokeratin expression in normal, hyperplastic and malignant epithelial cells from human prostate.
Studies were undertaken to define the expression of cytokeratins in normal, hyperplastic and malignant epithelial cells from human prostate. Cytokeratin (CK) polypeptides, separated by two-dimensional electrophoresis, were identified by immunoblotting with CK-specific monoclonal antibodies. CK polypeptides 5, 7, 8, 15, 18 and 19 were identified in fresh normal and hyperplastic prostate. Expression of CK 15 has not been previously reported in human prostate. Analysis of central and peripheral zone tissues from human prostate did not reveal qualitative differences in CK expression between these areas. Epithelial cells harvested from fresh BPH tissue by percoll gradient centrifugation and propagated in vitro using selective culture techniques showed alterations in CK expression compared to intact human prostate. Specifically, CKs 6, 14, 16 and 17 were noted in cultured BPH epithelial cells but not fresh normal prostate or BPH tissue. Immunoblot analysis of the established prostate cancer cell lines PC3, DU145 and LNCAP showed expression of CKs 8 and 18 but not CKs 5, 7 and 15 which were observed in benign prostate. These studies further characterize CK expression in benign and malignant human prostate and provide insights which may be useful in differentiating normal, hyperplastic and malignant epithelial cells in the human prostate gland.


1236 C/T and 3435 C/T polymorphisms of the ABCB1 gene in Mexican breast cancer patients.
MDR1, which is encoded by the ABCB1 gene, is involved in multidrug resistance (hydrophobic), as well as the elimination of xenotoxic agents. The association between ABCB1 gene polymorphisms and breast cancer risk in different populations has been described previously; however, the results have been inconclusive. In this study, we examined the association between polymorphisms 3435 C/T and 1236 C/T in the ABCB1 gene and breast cancer development in Mexican women according to their menopausal status and molecular classification. Molecular subtypes as well as allele and genotype frequencies were analyzed. A total of 248 women with initial breast cancer diagnosis and 180 ethnically matched, healthy, unrelated individuals were enrolled. Polymerase chain reaction-restriction fragment length polymorphism was performed to detect polymorphisms 3435 C/T and 1236 C/T in the ABCB1 gene. Premenopausal T allele carriers of the 3435 C/T polymorphism showed a 2-fold increased risk of breast cancer with respect to the reference and postmenopausal groups, as well as triple-negative expression regarding the luminal A/B molecular subrogated subtypes. In contrast, the CT genotype of the 1236 polymorphism was a protective factor against breast cancer. We conclude that the T allele carrier of the 3435 C/T polymorphism in the ABCB1 gene in combination with an estrogen receptor-negative status may be an important risk factor for breast cancer development in premenopausal women.


Clinical outcome of glass-fiber-reinforced crowns and fixed partial dentures: a three-year retrospective study.
Glass-fiber-reinforced resin composites for the fabrication of esthetic single crowns and fixed partial dentures have been developed recently. However, little data on the clinical outcome of such restorations have been published. The purpose of this retrospective study was to evaluate the clinical outcome of crowns and fixed partial dentures made from the glass-fiber-reinforced resin composite material Targis/Vectris. Between 1995 and 1997, 39 patients were treated with 67 single crowns and 83 fixed partial dentures. Twenty-five crowns and 28 fixed partial dentures were cemented with temporary cement, while 42 crowns and 55 fixed partial dentures were cemented with either zinc-phosphate cement or glass-ionomer cement. In September 2001, 57 restorations in 17 patients were still in function. The three-year survival rate calculated according to the Kaplan-Meier method was 58.8% for all restorations. However, after this time, it was 37.5% for crowns and 59.9% for fixed partial dentures cemented with temporary cement, and 55.1% for crowns and 67.9% for fixed partial dentures cemented with zinc phosphate or glass ionomer. The differences between the restoration groups were not statistically significant. Glass-fiber-reinforced crowns and fixed partial dentures made with the glass-fiber-reinforced resin composite material Targis/Vectris showed a lower survival rate than that published for metal-ceramic crowns and fixed partial dentures. Therefore, the use of this material for the fabrication of single crowns or fixed partial dentures as permanent restorations cannot be recommended.


Effectiveness and Safety in Remote Monitoring of Patients with Pacemakers Five Years after an Implant: The Poniente Study.
Health-related quality of life (HRQoL) and functional capacity values immediately after pacemaker (PM) implantation have been well established; however, not much has been known about its long-term effects. The present study compared the long-term effectiveness and safety of remote monitoring plus a clinic visit versus clinic visits alone during follow-up of adults implanted with PMs. This study was a single-centre, controlled, non-randomised, non-blinded clinical trial. Data were collected pre-implantation and after 60 months. The patients in the PONIENTE study were assigned to two different groups: remote monitoring (RM) and conventional monitoring (CM). The EuroQol-5D (EQ-5D) questionnaire was used to assess HRQoL and Duke Activity Status Index was used for the functional capacity. After five years, 55 patients completed the study (RM = 21; CM = 34). EuroQol-5D and functional capacity values were improved; however, significant differences were observed only in the EQ5D visual analogue scale (p < 0.001). Remote monitoring was equally feasible, reliable, safe, and clinically useful as CM. The frequencies of rehospitalisations and emergency visits did not differ between the groups. RM was found to be safe and effective in early detection and treatment of medical- and device-related events and in reducing hospital visits. Improved HRQoL was described not only immediately after PM implantation but also extended over a long time.


Association of radiomic imaging features and gene expression profile as prognostic factors in pancreatic ductal adenocarcinoma.
In this study, we investigated whether radiomic features of CT image data can accurately predict HMGA2 and C-MYC gene expression status and identify the patient survival time using a machine learning approach in pancreatic ductal adenocarcinoma (PDAC). A cohort of 111 patients with PDAC was enrolled in our study. Radiomic features were extracted using conventional (shape and texture analysis) and deep learning approaches following to segmentation of preoperative CT data. To predict patient survival time, significant radiomic features were identified using a log-rank test. After surgical resection, level of HMGA2 and C-MYC gene expressions of PDAC tumor regions were classified using a support vector machines method. The model was evaluated in terms of accuracy, sensitivity, specificity, and area under the curve (AUC). Besides, inter-reader reliability analysis was used to demonstrate the robustness of the proposed features. The identified features consistently achieved good performance in survival prediction and classification of gene expression status, on images segmented by different radiologists. Using CT data from 111 patients, six features in the segmented region of images were highly correlated with survival time. Using extracted deep features of excised lesions from 47 patients, we observed an average AUC score of 0.90 with an accuracy of 95% in C-MYC prediction (sensitivity: 92% and specificity: 98%). In HGMA2 group, using shape features, the average AUC score was measured as 0.91 with an accuracy of 88% (sensitivity: 89% and specificity: 88%). In conclusion, the radiomic features of CT image can accurately predict the expression status of HMGA2 and C-MYC genes and identify the survival time of PDAC patients.


Performance of endosonography-guided fine needle aspiration and biopsy in the diagnosis of pancreatic cystic lesions.
Preoperative diagnosis of cystic lesions of the pancreas remains difficult despite improvement in imaging modalities and cystic fluid analysis. The aim of our study was to assess the performance of endoscopic ultrasonography (EUS) and EUS-guided fine needle aspiration (FNA) in the diagnosis of pancreatic cystic lesions. Data from a series of 127 consecutive patients with pancreatic cystic lesions were prospectively studied. EUS and EUS-guided FNA were performed in all patients, and cystic material was used for cytological and histological analysis as well as for biochemical and tumor markers analysis. Performance of EUS diagnosis, biochemical and tumor markers, and FNA diagnosis were compared with the final histological diagnosis obtained at surgery or postmortem examination. Sixty-seven patients underwent surgery and therefore constituted our study group. EUS provided a tentative diagnosis in 113 cases (89%). Cytohistological FNA provided a diagnosis in 98 cases (77%). When the results of EUS and EUS-guided FNA were compared with the final diagnosis (67 cases), EUS correctly identified 49 cases (73%), whereas FNA correctly identified 65 cases (97%). Sensitivity, specificity, positive predictive value, and negative predictive value of EUS and EUS-guided FNA to indicate whether a lesion needed further surgery were 71% and 97%, 30% and 100%, 49% and 100%, and 40% and 95%, respectively. Carbohydrate antigen 19-9 > 50,000 U/ml had a 15% sensitivity and a 81% specificity to distinguish mucinous cysts from other cystic lesions, whereas it had a 86% sensitivity and a 85% specificity to distinguish cystadenocarcinoma from other cystic lesions. EUS-guided FNA is a valuable tool in the preoperative diagnostic assessment of pancreatic cystic lesions.


Anti-diabetic and anti-oxidative activity of fixed oil extracted from Ocimum sanctum L. leaves in diabetic rats.
Ocimum sanctum L. (OS) leaves have been shown to exert diverse potential benefits in a variety of stress conditions. The present study was conducted to elucidate the effects of the fixed oil extracted from OS leaves on the blood glucose levels and serum lipid profile of streptozotocin-induced diabetic rats. In addition, the anti-oxidative activity of OS leaves to protect various organs including the liver, kidney and heart was investigated. The fixed oil of the OS leaves was extracted using hexane, and the various fatty acid contents of the oil were determined using gas chromatography-mass spectrometry. Male Wistar rats were allocated into three groups (n=7 per group): Normal control rats, diabetic rats and diabetic rats fed daily with the fixed oil for three weeks. The results showed that α-linolenic acid was the primary fatty acid contained in the fixed oil of OS. After 3 weeks of diabetic induction, the rats exhibited increased blood glucose levels and serum lipid profile, in addition to elevated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatine kinase MB subunit (CK-MB), creatinine and blood urea nitrogen (BUN). The fixed oil significantly decreased the elevated levels of blood glucose, the serum lipid profile and the levels of serum creatinine and BUN (P<0.001), without exerting significant effects on the elevated serum levels of AST, ALT, LDH and CK-MB. Furthermore, the fixed oil increased the diabetically-reduced levels of serum insulin and decreased the rat kidney weight. Fixed oil suppressed the elevated thiobarbituric acid reactive substances (TBARS) level and increased the activity of various antioxidative enzymes in the rat renal tissue. By contrast, the fixed oil had no effect on the elevated TBARS level and the inhibited activity of the antioxidative enzymes in the rat liver and cardiac tissues. Histopathological results indicated that the fixed oil preserved the renal tissue against oxidative stress in diabetes. In summary, the results of the present study suggest that the fixed oil extracted from OS leaves exerted anti-hyperglycemic, anti-hyperlipidemic and free radical scavenging effects in diabetic rats, thus providing renal protection against diabetes. The α-linolenic acid contained in the fixed oil may be responsible for these effects.


The evolution of plasticity of dauer larva developmental arrest in the nematode Caenorhabditis elegans.
Organisms can end up in unfavourable conditions and to survive this they have evolved various strategies. Some organisms, including nematodes, survive unfavourable conditions by undergoing developmental arrest. The model nematode Caenorhabditis elegans has a developmental choice between two larval forms, and it chooses to develop into the arrested dauer larva form in unfavourable conditions (specifically, a lack of food and high population density, indicated by the concentration of a pheromone). Wild C. elegans isolates vary extensively in their dauer larva arrest phenotypes, and this prompts the question of what selective pressures maintain such phenotypic diversity? To investigate this we grew C. elegans in four different environments, consisting of different combinations of cues that can induce dauer larva development: two combinations of food concentration (high and low) in the presence or absence of a dauer larva-inducing pheromone. Five generations of artificial selection of dauer larvae resulted in an overall increase in dauer larva formation in most selection regimes. The presence of pheromone in the environment selected for twice the number of dauer larvae, compared with environments not containing pheromone. Further, only a high food concentration environment containing pheromone increased the plasticity of dauer larva formation. These evolutionary responses also affected the timing of the worms' reproduction. Overall, these results give an insight into the environments that can select for different plasticities of C. elegans dauer larva arrest phenotypes, suggesting that different combinations of environmental cues can select for the diversity of phenotypically plastic responses seen in C. elegans.


The prevalence and practices of caffeine use as an ergogenic aid in English professional soccer.
The ergogenic properties of caffeine are well established, with evidence supporting beneficial effects for physical and technical elements of performance required for successful soccer match play. Despite this, recommended caffeine practices for professional soccer have not been established. Therefore, the present study aimed to evaluate the use and behaviours surrounding caffeine use in elite English soccer clubs. Representatives of 36 clubs from the top four tiers of English professional football (40%) completed an online survey that sought to determine if, when, how and why caffeine was prescribed to players as a means of improving sports performance. Of the clubs sampled, 97% indicated that caffeine is provided to players as a means of improving performance. Caffeine is most commonly administered prior to (> 94%) and during a game (> 48%), with frequency uninfluenced by time of matches. There was a broad range and lack of consistency in the timing, dose and mode of caffeine administration, but doses were typically low. Evidence from the present study indicate a translational gap between science and practice, highlighting a need for future work to better understand how caffeine consumption can be optimised with respect to the specific demands and constraints in professional soccer.


Reduction of hepatic glucose production as a therapeutic target in the treatment of diabetes.
There has been an alarming increase in the population diagnosed with diabetes worldwide. Although there is an ongoing debate as to the role of liver in the pathogenesis of diabetes, reduction of hepatic glucose production has been targeted as a strategy for diabetes treatment. Indeed, reduction of hepatic glucose production can be achieved through modulation of both hepatic and extra-hepatic targets. This review describes the role of the liver in the control of glucose homeostasis. Gluconeogenesis and glycogenolysis are pathways for glucose production, whereas glycolysis and glycogenesis are pathways for glucose utilization/storage. At the biochemical and molecular level, the metabolic and regulatory enzymes integrate hormonal and nutritional signals and regulate glucose flux in the liver. Modulating either activities of or gene expression of these metabolic enzymes can control hepatic glucose production. Dysfunction of one or several enzyme(s) due to insulin deficiency or resistance results in increases in fluxes of glycogenolysis and gluconeogenesis and/or decreases in fluxes of glycolysis and glycogenesis, which thereby lead to glucose generation exceeding glucose consumption/disposal, as well as dysregulation of lipid metabolism. Activation of enzymes that promote glucose utilization/storage and/or inhibition of enzymes that reduce glucose generation achieve reduction of hepatic glucose production, and hence lower levels of plasma glucose in diabetes. This is also beneficial for the correction of dyslipidemia. Therefore, many enzymes are viable therapeutic targets for diabetes.


Red blood cell and plasma distribution in SHR cremaster muscle microvessels.
Distribution of blood in the cremaster muscle microvasculature of spontaneously hypertensive (SHR) and Wistar-Kyoto normotensive (WKY) rats was determined by fluorescence videomicroscopy and densitometry. Bolus intra-arterial injections of fluorescein isothiocyanate (FITC)-dextran or dichlorotriazenylaminofluorescein-treated red blood cells (DTAF-RBC) produced time-concentration curves in the series-coupled segments. WKY and SHR mean arterial pressure averaged 89 +/- 6 and 125 +/- 5 mmHg, respectively. Arteriolar diameters were not different between the two groups. The smallest SHR venules (V4) and larger diameters than those of WKY (P less than 0.05), whereas V3 and V2 were not different. WKY diameters for V1 were significantly greater than those of SHR. The mean transit (t) and appearance times (ta) of both indicators were equal at the largest arteriole (A1) in both groups. In A2, A3, and A4 the t for both indicators were generally lower in SHR than in WKY rats but not significantly. The venular t were significantly greater in SHR than in WKY rats with DTAF-RBC consistently less than FITC-dextran values. The greater cross-sectional area of V4 in SHR than WKY rats would reduce flow velocity and elevate t. Dispersion (ta/t = 1.0 indicates no dispersion) of both indicators was less in WKY than in SHR. The ratios for both groups increased at V4 because ta increased more than t. Shorter circuits for red blood cells than for plasma and less plasma skimming exists in WKY than in SHR.


SIRT1 - a new mammalian substrate of nuclear autophagy.
Macroautophagic/autophagic degradation of nuclear components (or nuclear autophagy) is a poorly understood area in autophagy research. We previously reported the nuclear lamina protein LMNB1 (lamin B1) as a nuclear autophagy substrate in primary human cells, stimulating the investigation of nuclear autophagy in the mammalian system. We recently reported the sirtuin protein SIRT1 as a new selective substrate of nuclear autophagy in senescence and aging. Upon senescence of primary human cells, SIRT1 degradation is mediated by a direct nuclear SIRT1-LC3 interaction, followed by nucleus-to-cytoplasm shuttling of SIRT1 and autophagosome-lysosome degradation. In vivo, SIRT1 is downregulated by lysosomes in hematopoietic and immune organs upon natural aging in mice and in aged human T cells. Our study identified another substrate of nuclear autophagy and suggests a new strategy to promote SIRT1-mediated health benefits by suppressing its autophagic degradation.Abbreviations: HSPC: hematopoietic stem and progenitor cells; NAD+: nicotinamide adenine dinucleotide; SASP: senescence-associated secretory phenotype.


Ascher Syndrome: Acquired Double Lower Lip Combined With Epulis Fissuratum.
Ascher syndrome is characterized by double lip, blepharochalasis, and non-toxic goiter triad. This condition of unknown etiology usually occurs in patients younger than 20 years of age. In this case report, the authors present the diagnostic process of Ascher syndrome and surgical procedure of double lower lip in a female patient who applied to our clinic for prosthetic reasons. A 70-year old female patient was referred to our clinic with the chief complaint of incompatible dentures. The patient had been using the same complete dentures for 40 years. Extraoral clinical examination revealed protruded, bulged lower lip at rest and bilateral blepharochalasis, defined as atrophy of the upper eyelid skin. The upper lip and lower eyelids were not affected. Intraoral examination revealed complete edentation of both jaws and a double lower lip, an accessory fold of excess mucosae inside the vermillion border. Furthermore, epulis fissuratum was observed in the lower jaw. There was no family history of double lip. The patient stated that she had noticed another lip inside her lip for several years. As Ascher syndrome was suspected, the patient was referred to the endocrinology department for thyroid examination, and the non-toxic goiter diagnosis was confirmed. Surgical removal of the redundant tissues was necessary because of deficiencies in speech, chewing, and aesthetics. The excess tissues were excised by transverse ellipse incision technique. Dentists should be familiar with Ascher syndrome, as they may be the first physicians to encounter patients with double lip.


Effects of hydrocortisone sodium succinate on the electrophysiological properties of hypoxic rabbit papillary muscle.
Hydrocortisone sodium succinate (HSS) (50 mg/1) was tested as a protective agent against the effects of hypoxia on rabbit papillary muscle fibres. No effect of HSS was observed on the electrophysiological variables studied in aerobic condition. Hypoxia (65-75 mmHg, 8.66-10.0 kPa) had no effects on the resting potential, amplitude and overshoot, and shortened the action potential at 0 and -60 mV levels of repolarization. In the presence of HSS the resting potential, amplitude and overshoot were also unaffected by the hypoxic condition. However, when measured at -60 mV level the changes on the repolarization phase after 15 and 60 minutes of hypoxia were significantly reduced, 14% and 24% respectively (p less than 0.001) as compared with the values obtained in the series without HSS.


A note on the balance between random sampling and population size. (On the 30th anniversary of G. Malécot's paper).
Wright's model for the effects of random fluctuations in gene frequency in a population of fixed size is generalized to randomly fluctuating population size, and treated from the viewpoint of G. Malécot, using a martingale convergence theorem. The gene frequency approaches a limit, whose value depends on the actual realization, or history, of the process; that is, convergence is with probability one (or: almost surely) in statistical language. The limit does not necessarily represent a state of fixation of either allele; in particular, the limiting probability distribution is not necessarily trivial. For the special case of deterministically varying population size, a necessary and sufficient condition for such non-triviality is given.


Physiology of mammalian prevertebral ganglia.
When assessing the physiological role of the prevertebral ganglia, the following should be considered. Ganglion cells in these ganglia receive continuous excitatory synaptic input from peripheral sensory mechanoreceptors and central preganglionic neurons. The latter may result from sensory afferents projecting from the gastrointestinal tract to spinal preganglionic neurons or as a result of endogenous oscillator activity located in the spinal cord (18) or within the ganglion (45). Peripheral and spinal reflex pathways probably do not operate as separate, independent pathways. Rather, they must be considered functionally integrated in the prevertebral ganglia. Because of continuous synaptic input, the prevertebral neuron operates at some level of "neurogenic tone" that provides capabilities for bi-directional responses. An increase in spinal or peripheral synaptic input will be matched by a proportional increase in output firing. The functional effect of spatial summation of peripheral and central input is to prime the prevertebral neuron so that adjustments in its firing frequency can be made without the lag inherent in building up a response in a quiescent system. Not all prevertebral ganglia participate in reflex activity. It may be that reflex activity between prevertebral ganglia and visceral smooth muscle depends upon the nature of the mechanical activity of the muscle. For example, the mechanical activity of smooth muscle of the vas deferens is not characterized by spontaneous, myogenic, regularly occurring contractions. Mechanical activity in this organ occurs in bursts. There is no evidence to suggest that sensory fibers project from the wall of the vas deferens to the hypogastric plexus thereby forming a peripheral reflex arc. Contraction of th smooth muscle of the vas deferens is brought about by bursts of nerve impulses through simple relay connections in the pelvic plexus. Smooth muscles supplied by this ganglion have a direct line to the central nervous system and are secured by a "fail-safe" system. In contrast, the mechanical activity of the gastrointestinal system is characterized by slow spontaneous myogenic activity. Pattern formation of motor activity is achieved by reflex activity within the enteric plexuses. Extrinsic alteration of the nature and pattern of contraction of smooth muscle of the gastrointestinal tract would be best served by prevertebral ganglia that modulate and adjust motor function by continuous integration of convergent weak synaptic input emanating from the peripheral and central nervous systems. When a prevertebral neuron is part of a peripheral reflex loop, its physiological role may be to modulate end organ activity; when it does not participate in a peripheral reflex loop, its role may be to relay CNS input and initiate end organ activity.


Effect of applied potentials on the activity and growth of Thiobacillus ferrooxidans.
The effect of applied DC potentials both in the positive and negative range, on the activity and growth of Thiobacillus ferrooxidans, is discussed. In general, application of positive potentials up to +1000 mV in an acid bioleaching medium was found to be detrimental to bacterial activity, while the impression of negative potentials enhanced both their activity and growth through electrochemical regeneration of ferrous ions and an increase in the biomass. Ferrous-ferric ratios in a bioleaching medium could be monitored through Eh measurements.Among the base sulfide minerals such as pyrite, chalcopyrite, and sphalerite, sphalerite could be selectively bioleached if an impressed potential of -500 mV (SCE) could be maintained in the leaching medium. Electrochemical bioleaching tests carried out under an applied potential of -500 mV with sphalerite in the presence and absence of noble minerals such as pyrite and chalcopyrite indicated enhanced zinc dissolution with negligible copper and iron in solution. Probable mechanisms and advantages of the electrochemical bioleaching process developed in the laboratory are outlined.


[The detection of the sleep apnea syndrome in a population of professional drivers].
The aim of this study was to reduce the risk of traffic accidents related to obstructive sleep apnea syndrome (OSAS) by means its detection and treatment in a group of 100 commercial drivers from Seville. Besides, to analyze which clinical findings could suggest OSAS. By means a questionnaire we selected subjects without (questionnaire score < 10 points) and with (questionnaire score > or = 10) clinical history of OSAS. In cases with score > or = 10, an overnight home polygraphy was carried out (Apnoescreen II, Jaeger), with measurement of oronasal airflow, chest and abdominal movements, oxygen saturation, electrocardiogram, body position and actimetry. We performed a manual analysis of recordings, and polygraphy was considered to be positive for OSAS if both AHI (respiratory events Index) or DI (desaturation Index) were > or = 10. In these positive cases, overnight conventional polysomnography was carried out (SleepLab, Jaeger), with therapeutic tests with CPAP if OSAS was diagnosed (AHI > or = 10). Average age and BMI (Body Mass Index) were 41.5 +/- 0.9 years and 28.2 +/- 0.4 kg/m2. Questionnaire was positive in 59 subjects, in 35 of whom home polysomnography was done. We did not find differences in age, BMI, neck circumference or symptomatology among these 35 drivers and the 24 remaining subjects in whom home polygraphy was not performed. Home polygraphy was positive in 10 subjects and negative In 25. Drivers in first group were older, heavier and complaint more frequently about snoring, sleep apnea and daytime sleepiness. We did not find differences in neck circumference, waist/hip ratio nor alcohol consumption between both groups. From the group with positive polygraphy, we performed conventional polysomnography in 8 cases and OSAS was diagnosed in 5 (in all cases, treatment with CPAP was started). This study does not provide data about prevalence, but it seems that the percentage of our drivers with OSAS could be lightly higher than the prevalence in general population. These subjects seem to be older, heavier and complaint more frequently about snoring, sleep apnea and daytime sleepiness.


Polycystin: in vitro synthesis, in vivo tissue expression, and subcellular localization identifies a large membrane-associated protein.
The primary structure of polycystin predicts a large integral membrane protein with multiple cell recognition motifs, but its function remains unknown. Insight into polycystin's normal function and its role in the development of autosomal dominant polycystic kidney disease (PKD1) requires the assembly of an extensive collection of molecular reagents to examine its expression and create model systems for functional studies. Development of these crucial reagents has been complicated due to the presence of transcriptionally active homologous loci. We have assembled the authentic full-length PKD1 cDNA and demonstrated expression of polycystin in vitro. Polyclonal antibodies directed against distinct extra- and intracellular domains specifically immunoprecipitated in vitro translated polycystin. The panel of antibodies was used to determine localization of polycystin in renal epithelial and endothelial cell lines and tissues of fetal, adult, and cystic origins. In normal adult kidney and maturing fetal nephrons, polycystin expression was confined to epithelial cells of the distal nephron and vascular endothelial cells. Expression in the proximal nephron was only observed after injury-induced cell proliferation. Polycystin expression was confined to ductal epithelium in liver, pancreas, and breast, and restricted to astrocytes in normal brain. We report clear evidence for the membrane localization of polycystin by both tissue sections and by confocal microscopy in cultured renal and endothelial cells. Interestingly, when cultured cells made cell-cell contact, polycystin was localized to the lateral membranes of cells in contact. These data suggest that polycystin is likely to have a widespread role in epithelial cell differentiation and maturation and in cell-cell interactions.


Optimum domestic processing and cooking methods for reducing the polyphenolic (antinutrient) content of pigeon peas.
Four high yielding and early maturing cultivars of pigeon pea (Cajanus cajan) namely UPAS-120, ICPL-87, ICPL-151 and, especially, Manak, contained significant amounts of polyphenols (1075 to 1328 mg/100g), which may limit their utilization. The effectiveness of soaking (6, 12 and 18 h, 30 degrees C), soaking and dehulling, ordinary cooking, pressure cooking and germination (24, 36, 48 h, 30 degrees C) in reducing the levels of polyphenols was investigated. A decrease in the polyphenolic contents varying from 4 to 26 percent in different pigeon pea cultivar was achieved. Pressure cooking of soaked-dehulled seeds was found to be the most effective method, followed by sprouting for 48 h, ordinary cooking of soaked-dehulled seeds, and pressure cooking of soaked whole seeds followed by sprouting for 36 h.


The Target Residence Time of Antihistamines Determines Their Antagonism of the G Protein-Coupled Histamine H1 Receptor.
The pharmacodynamics of drug-candidates is often optimized by metrics that describe target binding (Kd or Ki value) or target modulation (IC50). However, these metrics are determined at equilibrium conditions, and consequently information regarding the onset and offset of target engagement and modulation is lost. Drug-target residence time is a measure for the lifetime of the drug-target complex, which has recently been receiving considerable interest, as target residence time is shown to have prognostic value for the in vivo efficacy of several drugs. In this study, we have investigated the relation between the increased residence time of antihistamines at the histamine H1 receptor (H1R) and the duration of effective target-inhibition by these antagonists. Hela cells, endogenously expressing low levels of the H1R, were incubated with a series of antihistamines and dissociation was initiated by washing away the unbound antihistamines. Using a calcium-sensitive fluorescent dye and a label free, dynamic mass redistribution based assay, functional recovery of the H1R responsiveness was measured by stimulating the cells with histamine over time, and the recovery was quantified as the receptor recovery time. Using these assays, we determined that the receptor recovery time for a set of antihistamines differed more than 40-fold and was highly correlated to their H1R residence times, as determined with competitive radioligand binding experiments to the H1R in a cell homogenate. Thus, the receptor recovery time is proposed as a cell-based and physiologically relevant metric for the lead optimization of G protein-coupled receptor antagonists, like the H1R antagonists. Both, label-free or real-time, classical signaling assays allow an efficient and physiologically relevant determination of kinetic properties of drug molecules.


Observation of a continuum-continuum interference hole in ultrafast dissociating core-excited molecules.
The femtosecond dissociation of HCl after core excitation has been studied through the resonant Auger decay. The spectra contain contributions from decay occurring at both "molecular" and "atomic" internuclear distances. We have observed a new interference mechanism in these spectra: An atomic spectral line develops into a negative spectral contribution, a "hole," when detuning the excitation energy from the maximum of the Cl2p(-1)sigma(*) resonance. Resonant x-ray scattering theory quantitatively explains this behavior as due to a novel destructive continuum-continuum interference between molecular and atomic contributions to the Auger decay.


H5N1 Vaccine-Elicited Memory B Cells Are Genetically Constrained by the IGHV Locus in the Recognition of a Neutralizing Epitope in the Hemagglutinin Stem.
Because of significant viral diversity, vaccines that elicit durable and broad protection against influenza have been elusive. Recent research has focused on the potential of highly conserved regions of the viral hemagglutinin (HA) as targets for broadly neutralizing Ab responses. Abs that bind the highly conserved stem or stalk of HA can be elicited by vaccination in humans and animal models and neutralize diverse influenza strains. However, the frequency and phenotype of HA stem-specific B cells in vivo remain unclear. In this article, we characterize HA stem-specific B cell responses following H5N1 vaccination and describe the re-expansion of a pre-existing population of memory B cells specific for stem epitopes. This population uses primarily, but not exclusively, IGHV1-69-based Igs for HA recognition. However, within some subjects, allelic polymorphism at the ighv1-69 locus can limit IGHV1-69 immunodominance and may reduce circulating frequencies of stem-reactive B cells in vivo. The accurate definition of allelic selection, recombination requirements, and ontogeny of neutralizing Ab responses to influenza will aid rational influenza vaccine design.


Muscle blood flow, hypoxia, and hypoperfusion.
Blood flow increases to exercising skeletal muscle, and this increase is driven primarily by vasodilation in the contracting muscles. When oxygen delivery to the contracting muscles is altered by changes in arterial oxygen content, the magnitude of the vasodilator response to exercise changes. It is augmented during hypoxia and blunted during hyperoxia. Because the magnitude of the increased vasodilation during hypoxic exercise tends to keep oxygen delivery to the contracting muscles constant, we have termed this phenomenon "compensatory vasodilation." In a series of studies, we have explored metabolic, endothelial, and neural mechanisms that might contribute to compensatory vasodilation. These include the contribution of vasodilating substances like nitric oxide (NO) and adenosine, along with altered interactions between sympathetic vasoconstriction and metabolic vasodilation. We have also compared the compensatory vasodilator responses to hypoxic exercise with those seen when oxygen delivery to contracting muscles is altered by acute reductions in perfusion pressure. A synthesis of our findings indicate that NO contributes to the compensatory dilator responses during both hypoxia and hypoperfusion, while adenosine appears to contribute only during hypoperfusion. During hypoxia, the NO-mediated component is linked to a β-adrenergic receptor mechanism during lower intensity exercise, while another source of NO is engaged at higher exercise intensities. There are also subtle interactions between α-adrenergic vasoconstriction and metabolic vasodilation that influence the responses to hypoxia, hyperoxia, and hypoperfusion. Together our findings emphasize both the tight linkage of oxygen demand and supply during exercise and the redundant nature of the vasomotor responses to contraction.


Automatic detection and morphological delineation of bacteriophages in electron microscopy images.
Automatic detection, recognition and geometric characterization of bacteriophages in electron microscopy images was the main objective of this work. A novel technique, combining phase congruency-based image enhancement, Hough transform-, Radon transform- and open active contours with free boundary conditions-based object detection was developed to detect and recognize the bacteriophages associated with infection and lysis of cyanobacteria Aphanizomenon flos-aquae. A random forest classifier designed to recognize phage capsids provided higher than 99% accuracy, while measurable phage tails were detected and associated with a correct capsid with 81.35% accuracy. Automatically derived morphometric measurements of phage capsids and tails exhibited lower variability than the ones obtained manually. The technique allows performing precise and accurate quantitative (e.g. abundance estimation) and qualitative (e.g. diversity and capsid size) measurements for studying the interactions between host population and different phages that infect the same host.


Genome-Wide Analysis Reveals the Potential Role of MYB Transcription Factors in Floral Scent Formation in Hedychium coronarium.
The MYB gene family is one of the largest groups of transcription factors (TFs) playing diverse roles in several biological processes. Hedychium coronarium (white ginger lily) is a renowned ornamental plant both in tropical and subtropical regions due to its flower shape and strong floral scent mainly composed of terpenes and benzenoids. However, there is no information available regarding the role of the MYB gene family in H. coronarium. In the current study, the MYB gene family was identified and extensively analyzed. The identified 253 HcMYB genes were unevenly mapped on 17 chromosomes at a different density. Promoter sequence analysis showed numerous phytohormones related to cis-regulatory elements. The majority of HcMYB genes contain two to three introns and motif composition analysis showed their functional conservation. Phylogenetic analysis revealed that HcMYBs could be classified into 15 distinct clades, and the segmental duplication events played an essential role in the expansion of the HcMYB gene family. Tissue-specific expression patterns of HcMYB genes displayed spatial and temporal expression. Furthermore, seven HcMYB (HcMYB7/8/75/79/145/238/248) were selected for further investigation. Through RT-qPCR, the response of candidates HcMYB genes toward jasmonic acid methyl ester (MeJA), abscisic acid (ABA), ethylene, and auxin was examined. Yeast one-hybrid (Y1H) assays revealed that candidate genes directly bind to the promoter of bottom structural volatile synthesis genes (HcTPS1, HcTPS3, HcTPS10, and HcBSMT2). Moreover, yeast two-hybrid (Y2H) assay showed that HcMYB7/8/75/145/248 interact with HcJAZ1 protein. In HcMYB7/8/79/145/248-silenced flowers, the floral volatile contents were decreased and downregulated the expression of key structural genes, suggesting that these genes might play crucial roles in floral scent formation in H. coronarium by regulating the expression of floral scent biosynthesis genes. Collectively, these findings indicate that HcMYB genes might be involved in the regulatory mechanism of terpenoids and benzenoid biosynthesis in H. coronarium.


Teaching of primary care in an internal medicine residency program.
A teaching module in ambulatory internal medicine has been added to a residency program in a large referral hospital. The effort was directed to structure a program of instruction reflecting common problems seen by primary care internists. Patients were screened by supervising staff to make the rotation an efficient learning experience. Experienced nurse clinicians were used to monitor patients with stabilized medical problems, thus freeing the housestaff for problems more suited to their level of training. The nurses also served as role models with which the housestaff could interact and provide continuity and accessibility of care to patients being followed up by transient physicians-in-training. Various teaching conferences based on traditional inpatient models were conducted. Self-assessment and peer review techniques were structured around monthly conferences in which medical records were assessed by each participant.


Early Experience with the Octopus Endovascular Strategy in the Management of Thoracoabdominal Aneurysms.
The Octopus endovascular strategy involves placement of multiple, stacked bifurcated grafts in the thoracic segment of a thoracoabdominal aneurysm (TAAA) to facilitate deployment of multiple parallel covered stents for visceral perfusion. This study aimed to review early outcomes of the Octopus TAAA repair strategy at a tertiary, high-volume referral center. All patients who underwent this Octopus procedure from 2015 to 2018 were reviewed from a prospectively collected single-institution registry. Demographics, comorbidities, and aneurysm anatomy including side and extent, perioperative data including blood loss, length of procedure length of stay, morbidity, and mortality up to 3 years after the procedure were elevated. A total of 21 patients (48% female, age 72.9 years) underwent the Octopus procedure over the study period. Mean TAAA diameter was 6.7 cm, with 14% dissection related and 86% degenerative TAAA. All patients had been turned down for open repair and 3 (14%) were performed urgently of which 2 were ruptures. TAAA extent was 9% type 2, 62% type 3, and 29% type 4. A mean of 3.04 branches were revascularized per patient, with the superior mesenteric artery (SMA) (90%) perfused through its own limb, and both renals usually reconstructed in parallel graft fashion (left 90%, right 85.7%) with the distal abdominal extension through one of the limbs. Mean operative time was 8 hr, fluoroscopy time 164 min, contrast 182 mL, and blood loss 807 mL. We staged the thoracic and juxtavisceral portions of the cases in 24% of patients. 90% of cases were able to be completed with exclusion of the TAAA and all planned visceral branches cannulated. Perioperative complications included paraplegia (19%) (13.3% permanent, 26.6% temporary), acute kidney injury (24%), prolonged ventilation (19%), myocardial infarction (4.9%), and ischemic bowel (4.8%). Median follow-up was 13.5 months (range 1-26 months). At latest follow-up, type I endoleak rate was 9.5%, with all being treated with proximal cuffs. Other second interventions included restenting of a right renal, angioplasty of an iliac limb kink, and type 2 endoleak coiling. Primary patency of visceral branches was 93.8% at latest follow-up (celiac 100%, SMA 94.7%, right renal 88.9%, left renal 94.8%). In-hospital all-cause mortality rate was 14.2%, with 30-day survival being 90.5%, 6-month survival was 88.3%, one-year survival 71.4%, and 3-year survival was 52.1%. The Octopus procedure is a high-risk option for urgent or emergent endovascular TAAA repair with off-the-shelf devices in patients who are not candidates for open repair.


Metal-Free Radical Haloazidation of Benzene-Tethered 1,7-Enynes Leading to Polyfunctionalized 3,4-Dihydroquinolin-2(1H)-ones.
A new cascade three-component haloazidation of benzene-tethered 1,7-enynes for the formation of biologically interesting azidylated 3,4-dihydroquinolin-2(1H)-ones has been achieved under mild and metal-free conditions using TMSN3 as a N3 source and NIS (or NBS or NCS) as a halogen source. The reaction pathway involves in situ-generated azidyl radical-triggered α,β-conjugated addition/6-exo-dig cyclization/radical coupling sequence, resulting in successive multiple bond-forming events, including carbon-nitrogen, carbon-carbon, and carbon-halogen bonds to rapidly construct complex heterocyclic molecules. Furthermore, the resulting products would be useful building blocks in the discovery of lead compounds and other biologically interesting N3-containing heterocycles.


Acute administration of high doses of taurine does not substantially improve high-intensity running performance and the effect on maximal accumulated oxygen deficit is unclear.
The aim of the present study was to investigate the effects of acute administration of taurine overload on time to exhaustion (TTE) of high-intensity running performance and alternative maximal accumulated oxygen deficit (MAODALT). The study design was a randomized, placebo-controlled, crossover design. Seventeen healthy male volunteers (age: 25 ± 6 years; maximal oxygen uptake: 50.5 ± 7.6 mL·kg(-1)·min(-1)) performed an incremental treadmill-running test until voluntary exhaustion to determine maximal oxygen uptake and exercise intensity at maximal oxygen uptake. Subsequently, participants completed randomly 2 bouts of supramaximal treadmill-running at 110% exercise intensity at maximal oxygen uptake until exhaustion (placebo (6 g dextrose) or taurine (6 g) supplementation), separated by 1 week. MAODALT was determined using a single supramaximal effort by summating the contribution of the phosphagen and glycolytic pathways. When comparing the results of the supramaximal trials (i.e., placebo and taurine conditions) no differences were observed for high-intensity running TTE (237.70 ± 66.00 and 277.30 ± 40.64 s; p = 0.44) and MAODALT (55.77 ± 8.22 and 55.06 ± 7.89 mL·kg(-1); p = 0.61), which seem to indicate trivial and unclear differences using the magnitude-based inferences approach, respectively. In conclusion, acute 6 g taurine supplementation before exercise did not substantially improve high-intensity running performance and showed an unclear effect on MAODALT.


No evidence of conpopulation sperm precedence between allopatric populations of house mice.
Investigations into the evolution of reproductive barriers have traditionally focused on closely related species, and the prevalence of conspecific sperm precedence. The effectiveness of conspecific sperm precedence at limiting gene exchange between species suggests that gametic isolation is an important component of reproductive isolation. However, there is a paucity of tests for evidence of sperm precedence during the earlier stages of divergence, for example among isolated populations. Here, we sourced individuals from two allopatric populations of house mice (Mus domesticus) and performed competitive in vitro fertilisation assays to test for conpopulation sperm precedence specifically at the gametic level. We found that ova population origin did not influence the outcome of the sperm competitions, and thus provide no evidence of conpopulation or heteropopulation sperm precedence. Instead, we found that males from a population that had evolved under a high level of postcopulatory sexual selection consistently outcompeted males from a population that had evolved under a relatively lower level of postcopulatory sexual selection. We standardised the number of motile sperm of each competitor across the replicate assays. Our data therefore show that competitive fertilizing success was directly attributable to differences in sperm fertilizing competence.


Positive personality traits moderate persistent high alcohol consumption, determined by polygenic risk in U.S. military veterans: results from a 10-year, population-based, observational cohort study.
Understanding the interplay between psychosocial factors and polygenic risk scores (PRS) may help elucidate the biopsychosocial etiology of high alcohol consumption (HAC). This study examined the psychosocial moderators of HAC, determined by polygenic risk in a 10-year longitudinal study of US military veterans. We hypothesized that positive psychosocial traits (e.g. social support, personality traits, optimism, gratitude) may buffer risk of HAC in veterans with greater polygenic liability for alcohol consumption (AC). Data were analyzed from 1323 European-American US veterans who participated in the National Health and Resilience in Veterans Study, a 10-year, nationally representative longitudinal study of US military veterans. PRS reflecting genome-wide risk for AC (AUDIT-C) was derived from a Million Veteran Program genome-wide association study (N = 200 680). Among the total sample, 328 (weighted 24.8%) had persistent HAC, 131 (weighted 9.9%) had new-onset HAC, 44 (weighted 3.3%) had remitted HAC, and 820 (weighted 62.0%) had no/low AC over the 10-year study period. AUDIT-C PRS was positively associated with persistent HAC relative to no/low AC [relative risk ratio (RRR) = 1.43, 95% confidence interval (CI) = 1.23-1.67] and remitted HAC (RRR = 1.63, 95% CI = 1.07-2.50). Among veterans with higher AUDIT-C PRS, greater baseline levels of agreeableness and greater dispositional gratitude were inversely associated with persistent HAC. AUDIT-C PRS was prospectively associated with persistent HAC over a 10-year period, and agreeableness and dispositional gratitude moderated this association. Clinical interventions designed to target these modifiable psychological traits may help mitigate risk of persistent HAC in veterans with greater polygenic liability for persistent HAC.


Prevention of human papillomavirus (HPV)-related tumors in people living with human immunodeficiency virus (HIV).
In comparison to their HIV-negative counterparts, people living with HIV (PLWH) have a higher prevalence of human papillomavirus (HPV) infection in various anatomical sites coupled with increased HPV persistence, higher risk of HPV-related tumors, and faster disease progression. Areas covered: Gender-neutral prevention strategies for HPV-related cancers in PLWH discussed: ABC approach, HPV vaccination, antiretroviral treatment (ART), anal cancer screening, and smoking cessation. Gender specific strategies: cervical cancer screening reduces the incidence and mortality of cervical cancer and circumcision might reduce the risk of HPV infections in men. Expert commentary: HPV-related cancer incidence has not declined (e.g. cervical cancer) and has even increased (e.g. anal cancer) in the ART era, demanding an effective HPV prevention strategy. HPV vaccination should be introduced into national prevention programs worldwide immediately because current prophylactic vaccines are safe, tolerable, and immunogenic in PLWH. HPV vaccine efficacy trials in PLWH are essential to determine the most appropriate immunization schedule. The population most at risk of anal cancer is HIV-positive men who have sex with men, who are not protected by herd immunity if only the female population is vaccinated. Unvaccinated PLWH need enhanced surveillance for early detection of HPV-related cancers and their precursors.


Study the Effect of Calendula officinalis Extract Loaded on Zinc Oxide Nanoparticle Cream in Burn Wound Healing.
The skin, the body's largest organ, acts as a protective barrier against pathogens and environmental damage. Skin burns can result from heat, chemicals, friction, or electricity. Nanoscience has recently been utilized to create ointments and creams for burns. Zinc oxide nanoparticles are crucial due to their antimicrobial and antioxidant properties. In this study, a cream containing nanoparticles was loaded with calendula extract, and its ability to promote tissue healing was investigated in Wistar rats with skin burns. The zinc oxide nanoparticles were chemically synthesized and loaded with calendula extract. The morphology and physicochemical properties of the nanoparticles were confirmed by SEM, ZETA size, XRD, and FTIR assays. The MTT technique was employed to assess the cream's impact on fibroblast growth. The antimicrobial activity of the nanoparticles was investigated against Pseudomonas using the MIC method. Real-time PCR was used to determine the expression of the Bax and Bcl-2 genes in aeruginosa. The results showed that zinc oxide nanoparticles at high concentrations increased the proliferation of the fibroblast cells. Histopathological studies showed granulation and epithelialization of the tissue without any hemorrhage or tissue infection during the first days of treatment with this cream. The animal models treated with the cream showed an increase in Bcl-2 gene expression and a decrease in Bax expression. We concluded that zinc oxide nanoparticles loaded with calendula extract have a practical effect in healing burn wounds due to their unique antibacterial properties of zinc oxide nanoparticles and their anti-inflammatory and wound-healing effects. The synergistic effect of these two substances significantly improved the healing process. This newly developed cream can be introduced as a successful and viable treatment option in burn wounds.


Use of an internal standard in subtraction radiography to assess initial periodontal bone changes.
To assess the use of an internal reference when performing histogram analyses in digital subtraction images and to determine the ability of the method to detect initial bone lesions. Fifty-one Royal Air Force recruits had standardized vertical bitewing radiographs and clinical assessment of attachment level recorded annually over three years. Subtraction analyses of crestal bone changes at the mesial surface of the upper right first molar were compared with changes at the mesial surface of the same tooth. Changes over the periods from age 17 to 18 years and age 18 to 20 years were monitored using two subtraction procedures. Reproducibility studies revealed that Pearson correlations between duplicate measurements of the test site alone (16 crest; r = 0.74) were lower than those between duplicate measurements of changes where misalignment was controlled for, either as a difference (16 crest-16 tooth; r = 0.93) or ratio (16 crest/16 tooth; r = 0.93). We used the differences between the mean subtraction density for 16 crest and 16 tooth as our measure of change in bone density. For the duplicate measurements, the standard deviation of these differences was 3.9: a difference of +/- 7.8 was therefore taken as a threshold value for evidence of real change. Bone gain was noted between the ages of 17 and 18 years (16/21 subjects), but some early bone loss was seen between 18 and 20 years (12/21 subjects), with four subjects showing changes significantly greater than the method error. There were no associations between the clinical and radiographic observations. Use of a control site in subtraction radiography improves the reproducibility; such systems can detect small changes in alveolar bone which may assist in early diagnosis of the initial periodontal lesion which may precede observable clinical changes.


Vesicular trafficking of tyrosine kinase receptors and associated proteins in the regulation of signaling and vascular function.
Receptor tyrosine kinases (RTKs) play a pivotal role in the development and function of the cardiovascular system. Ligand-activated RTKs promote numerous downstream signal transduction pathways that lead to vascular permeability, as well as proliferation, migration, and differentiation of vascular endothelia and smooth muscle cells. Ligand binding also promotes internalization of the activated receptors either to downregulate the signaling via degradation of the ligand/receptor complex or to signal from endosomes. However, the outcomes of receptor internalization via clathrin-dependent or caveolar pathways and trafficking mechanisms are incompletely clarified in vascular systems. Activity modulation through endocytosis and vesicular trafficking significantly impacts downstream targets of RTKs such as endothelial nitric oxide synthase (eNOS) and VE-cadherin. RTKs and their associated targets are also transported to the nucleus, where they may directly impact nuclear signaling. Although the nuclear transport pathways are just beginning to be unraveled, it appears that endocytosis and vesicular trafficking are involved. In this review, we discuss the mechanisms by which activated RTKs and the downstream targets eNOS and VE-cadherin may be internalized and transported to various intracellular compartments. How localization and interacting proteins impact protein function and influence signaling is an important theme, as is the potential for modulating signaling through therapeutic targeting of activated receptors and components of the endocytic machinery.


Medical and surgical interventions for the treatment of usual-type vulval intraepithelial neoplasia.
Usual-type vulval intraepithelial neoplasia (uVIN) is a pre-cancerous condition of the vulval skin. Also known as high-grade VIN, VIN 2/3 or high-grade vulval squamous intraepithelial lesion (HSIL), uVIN is associated with high-risk subtype human papilloma virus (HPV) infection. The condition causes distressing vulval symptoms in the majority of affected women and may progress to vulval cancer, therefore is usually actively managed. There is no consensus on the optimal management of uVIN. High morbidity and recurrence rates associated with surgical treatments make less invasive treatments highly desirable. To determine which interventions are the most effective, safe and tolerable for treating women with uVIN. We searched the Cochrane Gynaecological Cancer Group Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL), Issue 8 2015, MEDLINE and EMBASE (up to 1 September 2015). We also searched registers of clinical trials, abstracts of scientific meetings, reference lists of included studies and contacted experts in the field. Randomised controlled trials (RCTs) that assessed medical and surgical interventions in women with uVIN. If no RCTs were available, we included non-randomised studies (NRSs) with concurrent comparison groups that controlled for baseline case mix in multivariate analysis. We used Cochrane methodology with two review authors independently extracting data and assessing risk of bias. Where possible, we synthesised data in meta-analyses using random-effects methods. Network meta-analysis was not possible due to insufficient data. We included six RCTs involving 327 women and five NRSs involving 648 women. The condition was variously named by investigators as uVIN, VIN2/3 or high-grade VIN. Five RCTs evaluated medical treatments (imiquimod, cidofovir, indole-3 carbinol), and six studies (one RCT and five NRSs) evaluated surgical treatments or photodynamic therapy. We judged two RCTs and four NRSs to be at a high or unclear risk of bias; we considered the others at relatively low risk of bias. Types of outcome measures reported in NRSs varied and we were unable to pool NRS data. Medical interventions: Topical imiquimod was more effective than placebo in achieving a response (complete or partial) to treatment at five to six months post-randomisation (three RCTs, 104 women; risk ratio (RR) 11.95, 95% confidence interval (CI) 3.21 to 44.51; high-quality evidence). At five to six months, a complete response occurred in 36/62 (58%) and 0/42 (0%) women in the imiquimod and placebo groups, respectively (RR 14.40, 95% CI 2.97 to 69.80). Moderate-quality evidence suggested that the complete response was sustained at one year (one RCT, nine complete responses out of 52 women (38%)) and beyond, particularly in women with smaller VIN lesions. Histologically confirmed complete response rates with imiquimod versus cidofovir at six months were 45% (41/91) and 46% (41/89), respectively (one RCT, 180 women; RR 1.00, 95% CI 0.73 to 1.37; moderate-quality evidence). Twelve-month data from this trial are awaited; however, interim findings suggested that complete responses were sustained at 12 months. Only one trial reported vulval cancer at one year (1/24 and 2/23 in imiquimod and placebo groups, respectively). Adverse events were more common with imiquimod than placebo and dose reductions occurred more frequently in the imiquimod group than in the placebo group (two RCTs, 83 women; RR 7.77, 95% CI 1.61 to 37.36; high-quality evidence). Headache, fatigue and discontinuation were slightly more common with imiquimod than cidofovir (moderate-quality evidence). Quality of life scores reported in one trial (52 women) were not significantly different for imiquimod and placebo. The evidence of effectiveness of topical treatments in immunosuppressed women was scant. There was insufficient evidence on other medical interventions. Surgical and other interventions: Low-quality evidence from the best included NRS indicated, when data were adjusted for confounders, that there was little difference in the risk of VIN recurrence between surgical excision and laser vaporisation. Recurrence occurred in 51% (37/70) of women overall, at a median of 14 months, and was more common in multifocal than unifocal lesions (66% versus 34%). Vulval cancer occurred in 11 women (15.1%) overall at a median of 71.5 months (9 to 259 months). The risk of vulval cancer did not differ significantly between excision and laser vaporisation in any of the NRSs; however, events were too few for robust findings. Alternative surgical procedures that might be as effective include Cavitron ultrasonic surgical aspiration (CUSA) and loop electrosurgical excision (LEEP) procedures, based on low- to very low-quality evidence, respectively. Very low-quality evidence also suggested that photodynamic therapy may be a useful treatment option.We found one ongoing RCT of medical treatment (imiquimod) compared with surgical treatment. Topical treatment (imiquimod or cidofovir) may effectively treat about half of uVIN cases after a 16-week course of treatment, but the evidence on whether this effect is sustained is limited. Factors predicting response to treatment are not clear, but small lesions may be more likely to respond. The relative risk of progression to vulval cancer is uncertain. However, imiquimod and cidofovir appear to be relatively well tolerated and may be favoured by some women over primary surgical treatment.There is currently no evidence on how medical treatment compares with surgical treatment. Women who undergo surgical treatment for uVIN have about a 50% chance of the condition recurring one year later, irrespective of whether treatment is by surgical excision or laser vaporisation. Multifocal uVIN lesions are at a higher risk of recurrence and progression, and pose greater therapeutic dilemmas than unifocal lesions. If occult cancer is suspected despite a biopsy diagnosis of uVIN, surgical excision remains the treatment of choice. If occult cancer is not a concern, treatment needs to be individualised to take into account the site and extent of disease, and a woman's preferences. Combined modalities may hold the key to optimal treatment of this complex disease.


Protein phosphatase 2C is responsible for VP-induced dephosphorylation of AQP2 serine 261.
Aquaporin 2 (AQP2) trafficking is regulated by phosphorylation and dephosphorylation of serine residues in the AQP2 COOH terminus. Vasopressin (VP) binding to its receptor (V2R) leads to a cascade of events that result in phosphorylation of serine 256 (S256), S264, and S269, but dephosphorylation of S261. To identify which phosphatase is responsible for VP-induced S261 dephosphorylation, we pretreated cells with different phosphatase inhibitors before VP stimulation. Sanguinarine, a specific protein phosphatase (PP) 2C inhibitor, but not inhibitors of PP1, PP2A (okadaic acid), or PP2B (cyclosporine), abolished VP-induced S261 dephosphorylation. However, sanguinarine and VP significantly increased phosphorylation of ERK, a kinase that can phosphorylate S261; inhibition of ERK by PD98059 partially decreased baseline S261 phosphorylation. These data support a role of ERK in S261 phosphorylation but suggest that, upon VP treatment, increased phosphatase activity overcomes the increase in ERK activity, resulting in overall dephosphorylation of S261. We also found that sanguinarine abolished VP-induced S261 dephosphorylation in cells expressing mutated AQP2 S256A, suggesting that the phosphorylation state of S261 is independent of S256. Sanguinarine alone did not induce AQP2 membrane trafficking, nor did it inhibit VP-induced AQP2 membrane accumulation in cells and kidney tissues, suggesting that S261 does not play an observable role in acute AQP2 membrane accumulation. In conclusion, PP2C activity is required for S261 AQP2 dephosphorylation upon VP stimulation, which occurs independently of S256 phosphorylation. Understanding the pathways involved in modulating PP2C will help elucidate the role of S261 in cellular events involving AQP2.


Penetrative and non-penetrative interaction between Laboulbeniales fungi and their arthropod hosts.
Laboulbeniales are a highly specialized group of fungi living only on arthropods. They have high host specificity and spend their entire life-cycle on an arthropod host. Taxonomic characters of Laboulbeniales are based on the architecture of the cells of the parenchymal thallus, i.e. the visible part of the fungus outside the host. The extent of the fungus spreading inside the host-the haustorium-remains largely unknown. The attachment to the arthropod host is fundamental to understand the fungus-animal interaction, but how this truly occurs is unclear. Recent evidences question the strictly parasitic life-style of Laboulbeniales. We used micro-computed tomography (µCT) and 3D reconstructions to visualize, for the first time, the complete structure of Laboulbeniales species in situ on their hosts. We compared the haustoriate species, Arthrorhynchus nycteribiae on an insect host to the non-haustoriate species, Rickia gigas on a millipede host. Our results confirm that some Laboulbeniales species are ectoparasitic and have a haustorial structure that penetrates the host's cuticle, while others are ectobionts and are only firmly attached to the host's cuticle without penetrating it. The presence and the morphology of the haustorium are important traits for Laboulbeniales evolution, and key factors for future understanding of host dependence and specificity.