[The effect of diet induced obesity on testicular tissue and serum oxidative stress parameters].
The aim of this study was to evaluate the effects of diet induced obesity on semen parameters and serum antioxidant enzyme levels. Six-week-old male rats were randomized into three groups are as follows: group 1 (n=10) received a control diet, group 2 (n=9) received a high-fat diet and group 3 (n=11) received high-fat diet plus anastrozole. At the completion of a 10-week period, testicular tissues were obtained and spermatogenesis was evaluated with Johnsen Score System. The normal Johnsen Score was accepted as >9.39. In addition, serum antioxidant enzyme levels, triglyceride, cholesterol, testosterone, luteinizing hormone (LH), follicle stimilating hormone (FSH) and estradiol levels were measured in serum. Body weight were significantly increased in mice fed with a high-fat diet compared to normal diet (P<.05). The mean triglyceride levels was 64.00±20.48 mg/dl, 98.89±27.80 mg/dl and 95.27±15.02 mg/dl in group 1, group 2 and group 3, respectively (P<.05). Male rats fed with a high-fat diet had significantly lower levels of testosterone compared with the control diet male rats (P=.005). Testicular pathology revealed that Johnsen Score System were 9.60±0.15, 8.72±1.81 and 9.29 in group 1, group 2 and group 3, respectively (P=.169). In addition serum nitric oxide (NO) levels were higher in group 2 and group 3 compared to group 1 (P<.05). As a result it may be concluded that obesity may induce oxidative stress and decrease testosterone levels. These changes may alter testicular functions and consequently it may be speculated that obesity can be important causative factor in the etiology of the male infertility.


Farmed wild boars exposed to Toxoplasma gondii and Trichinella spp.
The meat of wild boar (Sus scrofa L.) can be a source of human infections with zoonotic parasites Toxoplasma gondii and Trichinella spp. We screened 197 wild boar sera collected at slaughter from 25 Finnish farms in 2007-2008 for serological evidence of infections with these parasites. Using a commercial direct agglutination test at a serum dilution of 1:40, T. gondii-specific IgG antibodies were detected in 65 (33.0%) samples, on 14 (56.0%) farms. Females, animals older than 24 months, animals of small herds, and animals originating from south-western parts of Finland were more often T. gondii-seropositive than were males, younger animals, animals of larger herds, and animals originating from the north and east, respectively. Four (2.0%) of the sera, originating from three (12.0%) farms, tested Trichinella-seropositive with an in-house ELISA and a conservative cut-off for seropositivity. One farm had both T. gondii- and Trichinella-seropositive animals. Taken together, an infection source had been present on 16 (64.0%) farms, and 69 (35.0%) of the 197 farmed wild boars intended for human consumption had specific serological evidence of exposure to a zoonotic parasite.


Mother-child interactions: a comparison of a clinic-referred developmentally delayed group and two non-delayed groups.
The purpose of the present study was to compare a clinic-referred sample of developmentally delayed children to two groups of children who were not delayed: a clinic-referred sample and a nonclinic sample. Multiple outcome measures, including home observations of mother-child interactions, parental perceptions of child adjustment, and parental measures of depression and marital adjustment, were utilized. The results indicated that the delayed sample differed from the two nondelayed samples primarily in terms of emitting more maternal behavior, particularly the number of commands issued. A lower rate of compliance to total maternal commands also occurred for the delayed group. Finally, the clinic-referred delayed sample differed from the nonclinic non-delayed sample by perceiving their children as less well-adjusted. Implications of the results are discussed.


The impact of photobiomodulation therapy on the biology and behavior of head and neck squamous cell carcinomas cell lines.
Photobiomodulation therapy (PBMT) is an emerging therapeutic modality designed to prevent and treat chemotherapy-driven oral mucositis (OM). However, the response of tumor cells to the effects of PBMT remains poorly understood. Our study explores the effects of PBMT in head and neck squamous cell carcinoma (HNSCC) based on cellular proliferation, migration, and survival of tumor cells and its population of cancer stem cells (CSC). We explored the behavior of two HNSCC cell lines (HN6 and HN13) under two distinct conditions, a physiological growing condition (10% FBS), and under stress growing condition (2% FBS) prior to irradiation using diode laser (InGaAlP; MM Optics, São Carlos, SP, Brazil). Diode laser (660 nm) was applied with a power of 100 mW delivering a total energy per point of 0.24 J. MTT and wound healing test (scratch assay) were performed to evaluate, respectively, proliferation and migration of tumor cells. Clonogenic and spheres formation assays were also performed to evaluate the survival and percentage of CSC upon irradiation. Overall, we observed that PBMT does not exacerbate the behavior of HNSCC. We could only observe a decrease in cellular proliferation of one cell line (HN6) when cultured under nutritional stress conditions (p < .05). There were no significant differences between the control and the PBMT groups regarding cell migration, survival and the percentage of CSC. Collectively, our results suggest that in vitro administration of PBMT to HNSCC does not modify the behavior of tumor cells.


Acceptance of evidence-based reproductive health care among postgraduate specialist trainees in Nigeria.
To assess the perception and practice of evidence-based reproductive health care among Nigerian postgraduate specialist trainee doctors. A questionnaire survey of 326 Nigerian postgraduate specialist trainee doctors involved in reproductive health care was undertaken during training workshops on evidence-based medicine. Most respondents were familiar with evidence-based medicine (96.6%) and agreed that it would improve patient care (95.1%). Only 38.3% had ever received training on evidence-based medicine. Awareness and utilization of resources for evidence-based medicine were low; only 18.7% and 28.2% had ever used the Cochrane Library or the WHO Reproductive Health Library, respectively. Understanding of technical terms related to evidence-based medicine was poor. Lack of access to evidence was the most common constraint to the practice of evidence-based reproductive health care. Nigerian postgraduate specialist trainee doctors are positively disposed to evidence-based reproductive health care but their understanding of the principles of evidence-based practice is inadequate.


The effect of long-term use of cimetidine on the ultrastructure of gastric parietal cells in man.
The effect of long-term cimetidine therapy on the ultrastructure of human parietal cells was investigated in 2 subjects. It was found that after cimetidine therapy for 1 year: (i) the ultrastructural appearance of the parietal cells was normal, and (ii) the parietal cells responded normally to stimulation by pentagastrin. Evidence in favour of a membrane flow mechanism involved in acid secretion is presented.


Prostaglandin D2 (PGD2)--a potent coronary vasoconstrictor agent in the guinea pig isolated heart.
The action of prostaglandin D2 (PGD2) on myocardial force of contraction (MFC) and coronary vascular resistance (CVR) was studied in the isovolumetrically perfused guinea pig heart at constant driving frequency (180 beats/min). PGD2 (2 . 10(-9) - 1 . 10(-6) M) produced a concentration-dependent increase in the CVR while the MFC remained unchanged. The ED50 (50% of maximum response) of the coronary vasomotor action amounted to 4.3 . 10(-8) M PGD2. The results give evidence for a potent coronary vasoconstrictor activity of PGD2.


Nicotine-induced chondrogenic differentiation of human bone marrow stromal cells in vitro.
Nicotine has been reported that it has a dose-dependent effect on matrix mineralization by human bone marrow cells. However, there is no relevant research concerning on chondrogenic differentiation potential of bone marrow stromal stem cells (BMSCs) treated with nicotine in vitro. The aims of the study were to examine the effects of nicotine (0, 10(-7), 10(-6) and 10(-5) M) on the proliferation and chondrogenic differentiation of BMSCs from three healthy donors in vitro. BMSCs proliferation was analyzed by CCK8 assay and real-time polymerase chain reaction was used to assay the expression of type II collagen, aggrecan, type I collagen and type X collagen. The proteoglycan content was stained by Alcian blue, and the sulfated glycosaminoglycan (sGAG) content of BMSCs was quantified spectrofluorometrically using dimethylmethylene blue. The cell viability was not significantly impaired until up to a concentration of 10(-5) M nicotine. Nicotine promoted the proliferation and enhanced the expression of type II collagen at the level up to 10(-6) M (P < 0.05). The expression of aggrecan was reduced at the concentration of 10(-5) M nicotine at day 14 (P < 0.05), and there was no significant difference in aggrecan gene expression at 10(-7) and 10(-6) M nicotine levels compared to control group (n.s.). Also the fibroblastic and hypertrophic gene expressions were down-regulated in the chondrogenic medium with 10(-7)-10(-5) M nicotine (P < 0.05). It was implied that local application of nicotine at an appropriate concentration may be a promising approach for enhancing chondrogenic differentiation capacity of BMSCs in cell-based cartilage tissue engineering. Also these results indicate that nicotine maybe a potentially useful drug for the treatment of Osteoarthritis.


Correlation between middle cerebral artery peak systolic velocity and fetal hemoglobin after 2 previous intrauterine transfusions.
The middle cerebral artery peak systolic velocity (MCA-PSV) has been successfully used for timing the first 2 transfusions in fetuses at risk for anemia because of maternal red cell alloimmunization. The objective of this study was to assess whether the correlation between the MCA-PSV and fetal hemoglobin is maintained in fetuses that had undergone 2 previous intrauterine transfusions. Doppler measurement of MCA-PSV was performed before cordocentesis in 39 fetuses. The timing of the third transfusion was based on traditional criteria. The values of MCA-PSV and hemoglobin were expressed as multiples of the median (MoM). Anemia was defined as mild (hemoglobin <0.84 MoM for a given gestational age, moderate (hemoglobin <0.65 MoM), and severe (hemoglobin <0.55 MoM). Regression analysis was used to assess the correlation between the MCA-PSV MoM and fetal hemoglobin MoM. Gestational age at Doppler study ranged from 22 to 35 weeks. Six fetuses (15%) had normal hemoglobin concentration; 21 (53%) had mild anemia; 7 (20%) had moderate anemia; and 5 (12%) had severe anemia. There was a linear correlation between fetal hemoglobin (y) and the MCA-PSV (x): y = 1.185 - 0.341x. Previously, concerns have been expressed about the accuracy of Doppler prediction of anemia after previous transfusions. Our data suggest that there is a good correlation between the MCA-PSV and fetal hemoglobin in fetuses that have undergone 2 previous transfusions. Our findings expand the clinical situation in which Doppler can be used to monitor red cell alloimmunized pregnancies.


Exacerbation of cold restraint-induced gastric ulcer by GABA in mice.
To study the effect of GABA on cold restraint-induced gastric ulcer (CRGU) and its mechanism. CRGU was produced in adult male mice at 1 degree C in cold room under restraint for 1 h. The ulcer index indicated the severity of gastric mucosa lesion. Gastric mucin was determined with alcian blue dye. GABA 2 mumol (i.c.v.) accelerated CRGU, Bicuculline (Bic) 0.2 mumol (i.c.v.) did not affect CRGU, whereas Baclofen (Bac) 4 mumol (i.c.v.) exacerbated CRGU. Bic did not modify the exacerbation of CRGU by GABA and Bac. Atropine (Atr) 0.2 mg.kg-1 (s.c.) and Phentolamine (Phen) 2.5 mg.kg-1 inhibited CRGU, and abolished the exacerbation of CRGU by GABA. Cold restraint (CR) decreased the amount of the gastric mucin, but GABA 2 mumol had no effect on CR gastric mucin. While GABA-B receptor in brain was activated, exogenous GABA exacerbated CRGU via vagal and sympathetic nerves, bearing no relation to decrease of the gastric mucin or weakening of the gastric mucosal barrier.


A new case of Creutzfeldt-Jakob disease associated with human growth hormone therapy in New Zealand.
A clinically atypical, neuropathologically verified case of Creutzfeldt-Jakob disease is described in a 32-year-old New Zealand woman with idiopathic hypopituitarism who had been treated in late adolescence (1970 to 1973) with human growth hormone processed from pooled cadaveric pituitary glands.


Visualizing and quantitating sequence-dependent GPCR recycling.
Recent advances in direct imaging have given us a new appreciation of the spatial and temporal dynamics of membrane trafficking processes, and have allowed us to ask questions that were difficult to address with traditional methods. A relevant example of this is protein sorting in the endosome, which serves as the primary sorting station for proteins internalized from the cell surface. In this chapter, we discuss fluorescence imaging protocols to directly visualize and quantitate the recycling of G protein-coupled receptors (GPCRs)-a highly physiologically relevant family of signaling receptors-in real time in living cells. The protocols allow direct visualization and quantitation of both GPCR exit from the endosome and GPCR delivery to the cell surface. The methods may be extended to study the endolysosomal sorting of many proteins that undergoes endocytic cycling, and may be adapted to other organelles and systems where proteins are sorted.


In situ anterior vaginal wall sling formation with preservation of the endopelvic fascia for treatment of stress urinary incontinence.
The indications for sling procedures have evolved and encompass patients with either intrinsic sphincteric deficiency (ISD), anatomic incontinence or both. We have refined a technique that can be performed in a minimally invasive fashion with low attendant morbidity to provide a reproducible method of sling formation. Twenty patients with stress urinary incontinence underwent the in situ sling (ISS) with bone fixation. Subsequent evaluation at 24-29 months (mean = 26.2 months) revealed that 95% of patients were cured. No recurrent cystoceles, paravaginal defects or significant detrusor instability have been noted. Urinary retention appeared transiently in only 3 patients and resolved in under 3 weeks. We feel the in situ sling with bone fixation provides a safe and effective means of management for stress urinary incontinence. Furthermore, the reduced surgical dissection may minimize the incidence of postoperative ISD and recurrent paravaginal defects that may accompany more traditional needle suspension procedures.


Inflammatory bowel disease in mother or father and neonatal outcome.
Even a minor decrease in birthweight predisposes to adult disease. Inflammatory bowel disease (IBD) in the mother is a risk factor for low birthweight and preterm infants. This study investigated the effect of IBD in the mother or father, adjusting for confounders, on the newborn infant, with the focus on birthweight and pregnancy duration. A total of 10399 single-birth mother-infant pairs was prospectively enrolled within the ABIS project (All Babies In Southeast Sweden). Outcome measures included birth week, preterm birth (<37 wk), birthweight, low birthweight (<2500 g), birth length, caesarean section and neonatal hospital care. Ulcerative colitis (UC) in the mother was associated with lower birthweight in the infant (adjusted difference: -330 g, adjusted 95% confidence interval: -509 to -150 g, p < 0.001), and with even lower birthweight when the mother was treated with Mesalazine or steroids. No decrease in birthweight was seen in infants whose mother suffered from Crohn's disease (CD) (adjusted difference: -65 g, adjusted 95% confidence interval: -354 to 224 g, p > 0.05). Maternal UC or CD did not affect the pregnancy duration. The neonatal outcome of infants whose father suffered from UC and CD did not differ from the control group. UC in the mother affects the birthweight of the infant, which may predispose to future disease in the infant. Most women and men with UC and CD can, however, expect a healthy child with neither preterm birth nor low birthweight.


Use of flexible CO₂ laser fiber in microsurgery for vestibular schwannoma via the middle cranial fossa approach.
The aim of this study was to analyze the results of microsurgery in vestibular schwannomas (VS) with assistance of a flexible CO(2) laser fiber (Omniguide(®)) using the middle cranial fossa (MCF) approach. For that purpose we performed a prospective non-randomized clinical trial. In 20 consecutive patients suffering from VS and elected for microsurgery via the MCF approach, tumor resection was performed with the aid of the flexible CO(2) laser ("laser group", LG). Twenty patients with similar tumor volume and pre-operative hearing status out of a cohort of 76 patients previously treated by the same surgeon without laser were used as comparison group ("conventional group", CG) (matched-pair-technique). Facial weakness (House-Brackmann (HB) 2-4) was seen in early postoperative (p.o.) days in six patients in each group and all recovered completely by 3 months p.o., except one patient with HB 2 in CG. Facial nerve preservation rate (HB 1 + 2) was 100% in both groups. Hearing preservation rate (Gardner/Robertson class 1 + 2 or AAO-HNS A + B, pre- and postoperatively) was 72% in LG and 82% in CG, without significant difference. Overall time from incision to skin suture was 157 min (SD 55.9) in CG and 160 min (SD 39.7) in LG. Tumor preparation time was 23.2 min (SD 19.7) in CG and 36.1 min (SD 33.8) in LG. The use of a handheld flexible CO(2) laser fiber in VS-microsurgery is safe and subjectively facilitates tumor resection especially in "difficult" (e.g., highly vascularized) tumors. However, in this limited prospective trial the excellent functional outcome following conventional microsurgery could not be further improved, nor the surgical time reduced by means of the non-contact laser-tool. Focusing the use of the flexible CO(2) laser on "difficult" tumors may lead to different results in future.


Referral patterns and service utilization in a pediatric hospital-wide intimate partner violence program.
To describe the referral patterns and utilization of on-site intimate partner violence (IPV) services in both inpatient and outpatient settings at a large urban children's hospital. Retrospective review of case records from IPV victims referred to an on-site IPV counselor between September 2005 and February 2010. Descriptive statistics were used to examine IPV victim demographics, number of referrals per hospital department, referral source (type of staff member), time spent by IPV counselor for initial consultation, and services provided to IPV victims. A total of 453 unique referrals were made to the IPV counselor: 81% were identified by universal screening and 19% by risk-based screening. Thirty-six percent of IPV victims were referred from primary care clinics; 26% from inpatient units; 13% from outpatient subspecialty clinics; 12.5% from the emergency department; 5% from the Child Protection Program; and 4% were employee self-referrals. Social workers generated the most referrals (55%), followed by attending physicians (17%), residents (13%), nurses (7%), and other individuals (self-referrals) (4%). The median initial IPV intervention required 42 minutes. Supportive counseling and safety planning were the services most often utilized by IPV victims. IPV screening can be successfully integrated in both inpatient and outpatient settings by a multidisciplinary group of hospital staff. Most referrals were generated by universal screening outside of the primary care setting. IPV victims generally desired supportive counseling and safety planning over immediate housing relocation. Many IPV screening opportunities were missed by using verbal screening alone.


Evidence for linkage of postchallenge insulin levels with intestinal fatty acid-binding protein (FABP2) in Mexican-Americans.
Single genes with large effects may contribute to insulin resistance or influence susceptibility to non-insulin-dependent diabetes mellitus (NIDDM). In the Pima Indians, results from sib-pair analysis have suggested that a gene on chromosome 4q influences both fasting insulin levels and maximal insulin action. We conducted sib-pair and logarithm of odds (LOD)-score linkage analysis to seek evidence for linkage between genes influencing insulin levels and chromosome 4q loci. Analyses were conducted on nondiabetic individuals from 28 different families participating in the San Antonio Family Diabetes Study. All subjects received a 2-h oral glucose tolerance test. Fasting insulin levels were measured in 382 nondiabetic individuals, and 2-h insulin levels were measured in 366 individuals. Initial sib-pair linkage analysis revealed a possible association between 2-h post-glucose challenge insulin levels and the intestinal fatty acid-binding protein (FABP2) locus located in the region of chromosome 4q28-31 (P = 0.006). Subsequent sib-pair linkage analysis of 11 additional chromosome 4q markers supported this hypothesis. We next conducted segregation analyses to estimate allele frequencies and other model parameters for the putative locus influencing 2-h insulin levels. Results of LOD-score linkage analysis indicated possible linkage between the major gene described by the segregation model and FABP2. Using combined segregation and linkage analysis, we obtained a LOD-score of 2.80 at recombination frequency of 0.0 between FABP2 and the putative locus influencing 2-h insulin levels. The maximum likelihood estimate of the allele associated with low insulin levels was 0.21.(ABSTRACT TRUNCATED AT 250 WORDS)


Insights into the Mechanism of n-Hexane Reforming over a Single-Site Platinum Catalyst.
We demonstrate that the single-site catalyst Pt1/CeO2 greatly enhances the selectivity of cyclization and aromatization in the n-hexane reforming reaction. Specifically, the selectivity of single-site Pt1/CeO2 toward both cyclization and aromatization is above 86% at 350 °C. The turnover frequency of Pt1/CeO2 is 58.8 h-1 at 400 °C, which is close to that of Pt cluster/CeO2 (61.4 h-1) and much higher than that of Pt nanoparticle/CeO2 with Pt sizes of 2.5 and 7 nm. On the basis of the catalytic results for methylcyclopentane reforming, the dehydrocyclization and further aromatization of n-hexane are attributed to the prominent adsorption of ring intermediate products on the single-site Pt1/CeO2 catalysts. On the other side, with the multiple Pt adjacent active sites, the cluster and nanoparticle Pt/CeO2 samples favor the C-C bond cracking reaction. Ultimately, this in-depth study unravels the principles of hydrocarbon activation with different Pt sizes and represents a key step toward the rational design of new heterogeneous catalysts.


A correction in the CDM methodological tool for estimating methane emissions from solid waste disposal sites.
Solid waste disposal sites (SWDS) - especially landfills - are a significant source of methane, a greenhouse gas. Although having the potential to be captured and used as a fuel, most of the methane formed in SWDS is emitted to the atmosphere, mainly in developing countries. Methane emissions have to be estimated in national inventories. To help this task the Intergovernmental Panel on Climate Change (IPCC) has published three sets of guidelines. In addition, the Kyoto Protocol established the Clean Development Mechanism (CDM) to assist the developed countries to offset their own greenhouse gas emissions by assisting other countries to achieve sustainable development while reducing emissions. Based on methodologies provided by the IPCC regarding SWDS, the CDM Executive Board has issued a tool to be used by project developers for estimating baseline methane emissions in their project activities - on burning biogas from landfills or on preventing biomass to be landfilled and so avoiding methane emissions. Some inconsistencies in the first two IPCC guidelines have already been pointed out in an Annex of IPCC latest edition, although with hidden details. The CDM tool uses a model for methane estimation that takes on board parameters, factors and assumptions provided in the latest IPCC guidelines, while using in its core equation the one of the second IPCC edition with its shortcoming as well as allowing a misunderstanding of the time variable. Consequences of wrong ex-ante estimation of baseline emissions regarding CDM project activities can be of economical or environmental type. Example of the first type is the overestimation of 18% in an actual project on biogas from landfill in Brazil that harms its developers; of the second type, the overestimation of 35% in a project preventing municipal solid waste from being landfilled in China, which harms the environment, not for the project per se but for the undue generated carbon credits. In a simulated landfill - the same amount of waste for 20 years -, the error would be an overestimation of 25% if the CDM project activity starts from the very first year or an underestimation of 15% if it starts just after the landfill closure. Therefore, a correction in the tool to calculate emissions from landfills as adopted by the CDM Executive Board is needed. Moreover, in countries not using the latest IPCC guidelines, which provides clear formulas to prevent misunderstandings, inventory compilers can also benefit from this paper by having more accurate results in national GHG inventories related to solid waste disposal, especially when increasing amounts of waste are landfilled, which is the case of the developing countries.


Thermoluminescence studies of zinc borate (ZnB2 O4 ) phosphor doped with rare earths for dosimetric aspects.
A series of ZnB2 O4 phosphors doped with different concentrations of Eu and Dy (0.05 0.1, 0.2, 0.5, 1.0 mol%) and co-doped with Ce (1, 2, 5, 7, 10 mol%) respectively was prepared via the solid-state reaction technique and the thermoluminescence (TL) behaviour of gamma ray-irradiated samples was studied. The synthesized samples were irradiated with γ-rays for the dose range 0.03-1.20 kGy. The TL intensity variations with dose, dopant concentration, and the effect of co-doping were studied. The TL response curves for ZnB2 O4 :Eu3+ and ZnB2 O4 :Dy3+ , ZnB2 O4 :Eu3 ,Ce3+ and ZnB2 O4 :Dy3+ ,Ce3+ phosphor were observed. It was revealed that ZnB2 O4 :Eu3+ showed a linear TL behaviour for the dose 0.03-1.20 kGy and ZnB2 O4 :Dy3+ showed linearity for the gamma dose range 0.03-0.10 kGy. Furthermore, fading for all the samples was observed to be less than 10% for a storage period of 30 days. In addition to this, the trapping parameters, especially activation energies were evaluated using the Ilich method and the initial rise method. The activation energy values obtained from both methods were in complete agreement with each other.


Segmentation of arteries in MPRAGE images of the ventral medial prefrontal cortex.
A method for removing arteries that appear bright with intensities similar to white matter in Magnetized Prepared Rapid Gradient Echo images of the ventral medial prefrontal cortex is described. The Fast Marching method is used to generate a curve within the artery. Then, the largest connected component is selected to segment the artery which is used to mask the image. The surface reconstructed from the masked image yielded cortical thickness maps similar to those generated by manually pruning the arteries from surfaces reconstructed from the original image. The method may be useful in masking vasculature in other cortical regions.


Nitric oxide regulates AKT phosphorylation and nuclear translocation in cultured retinal cells.
Previous studies have shown that nitric oxide (NO) inhibits apoptosis of retinal neurons in culture through the canonical cyclic GMP/protein kinase G (PKG)-dependent pathway, but also involving multiple kinase pathways, such as phosphatidylinositol 3' kinase (PI3k) and AKT. NO and AKT exhibit survival-promoting properties and display important roles in both CNS development and plasticity. The purpose of this study was to evaluate the effects of exogenous NO, derived from the NO donor S-nitroso-N-acetylpenicillamin (SNAP), or endogenous NO, produced from l-arginine, on AKT phosphorylation in cultured chick retinal neurons. Our results demonstrate that SNAP or l-arginine enhances AKT phosphorylation on both serine-473 and threonine-308 residues in a concentration and time-dependent manner. This effect was mediated by the activation of soluble guanylyl cyclase and PKG, since it was blocked by the respective enzyme inhibitors ODQ or LY83583 and KT5823, as well as by transduction with shRNA lentiviruses coding PKGII shRNA, and mimicked by the respective enzyme activators YC-1 and 8-Bromo cyclic GMP, and also by the cyclic GMP phosphodiesterase inhibitor zaprinast. In addition, LY294002 or wortmannin suppressed the SNAP effect, indicating the involvement of phosphoinositide 3' kinase. Moreover, the mTOR inhibitor KU0063794 blocked SNAP-induced AKT phosphorylation at both residues, suggesting the participation of the mTORC2 complex in the process. Glutamate and NMDA also promoted AKT phosphorylation and a nitric oxide synthase inhibitor abrogated these effects, revealing a mechanism involving the activation of NMDA receptors and NO production. We have also found that SNAP and l-arginine induced AKT translocation into the nucleus of retinal neurons as well as other neuronal cell lines. SNAP also protects retinal cells from death induced by hydrogen peroxide and this effect was blocked by the phosphoinositide 3' kinase inhibitor LY294002. We therefore conclude that NO produced from endogenous or exogenous sources promotes AKT activation and its shuttling to the nucleus, probably participating in neuronal survival pathways important during CNS development.


Pudendal Neurolysis by Laparoscopy.
To show how pudendal neurolysis can be managed safely with a laparoscopic approach. Stepwise demonstration of the technique with narrated video footage. The pudendal nerve is formed from spinal roots at levels S2, S3, and S4. It passes through the pelvis parallel to the pudendal vein and artery. This nerve exits the pelvis between the sacrospinous and sacrotuberous ligaments then passes through Alcock's canal. It can be compressed and responsible for pain in the gluteal and perineal regions. After confirmation of the diagnosis by positive analgesic block with computed tomography infiltration of the pudendal nerve, surgical decompression may be considered. The usual access procedures are the transglutal and transischiorectal ways. This video shows a total laparoscopic approach for a right pudendal neurolysis. It is a step-by-step didactic video. This technique of decompression of the right pudendal nerve by laparoscopy by means of dissection of the ischiorectal fossa along the right internal obturator muscle, after visualization of the obturator vessels and identification of the pudendal nerve, allowed the section of the right sacrospinous ligament and complete removal with repositioning of the nerve in its path. The nerve was followed until it passed freely through Alcock's canal. The procedure went well and without complications, with clinical improvement on waking up. Pudendal nerve neurolysis by laparoscopic technique is a reproducible and safe method for treating pudendal neuralgia, allowing good visualization and dissection of the entire pelvis toward the ischiorectal fossa.


Localization of enkephalin- and neurotensin-like immunoreactivities in cat adrenal medulla.
Enkephalin (ENK)- and neurotensin (NT)-immunoreactivities (IR) were localized in cat adrenal medulla using immunocytochemistry. ENK was localized mainly in adrenaline cells, 70%-80% of which were heavily labelled. About 5%-10% of the noradrenaline cells were ENK-immunoreactive (IR). A dense network of ENK-IR nerves was localized among adrenaline cells. NT was localized only in the noradrenaline cells, 60%-70% of which were immunoreactive. NT- and ENK-IR were localized in separate noradrenaline cells. A dense network of NT-IR nerves was restricted among noradrenaline cells. The present findings confirm that ENK- and NT-like peptides are localized in separate adrenal medullary cells and demonstrate that also the preganglionic nerves are divided in subpopulations according to their neuropeptide content. These nerves have a selective localization among adrenaline and noradrenaline cells and may have physiological significance in the control of secretion of catecholamines and neuropeptides from adrenal medulla.


Growth performance, carcass and meat quality, bone strength, and immune response of broilers fed low-calcium diets supplemented with marine mineral complex and phytase.
Influence of marine mineral complex (CeltiCal) as a partial substitute for limestone on growth efficiency, carcass traits, meat quality, bone strength, calcium (Ca) retention, and immune response was investigated in broilers fed low-Ca diets with or without phytase (PHY) addition for a 35-d trial period. A total of 300 one-day-old Ross 308 straight-run broilers were randomly allocated to: T1 (positive control), recommended Ca levels + PHY; T2 (negative control), 0.2% below the recommended Ca levels + PHY; T3, 0.1% below the recommended Ca levels + 0.2% CeltiCal + PHY; T4, 0.2% below the recommended Ca levels + 0.4% CeltiCal + PHY; T5, 0.2% below the recommended Ca levels + 0.4% CeltiCal. PHY was added at 500 phytase units/kg diets. Each dietary treatment had 10 replications of 6 chicks each. Results revealed that production efficiency factor was greater for T4 compared to T2 and T5 during 22-35 d and for T1, T3, and T4 compared to T2 during 0 to 35 d (P < 0.05). Feed conversion ratio was lower for T3 and T4 compared to T2 and T5 during 0 to 35 d (P < 0.05). T4 had a greater (P < 0.05) dressing percentage than T2, which had a lighter (P < 0.01) small intestinal relative weight than all other treatments. Breast meat temperature at 15 min postmortem was highest for T1 and lowest for T3 (P < 0.001). Breast meat pH was greater for T1 compared to T5 at 15 min postmortem and for T3 compared to T4 at 24 h postmortem (P < 0.05). T5 had a lower breast meat redness than all other treatments at 15 min postmortem and then T1 and T3 at 24 h postmortem (P < 0.01). Tibia and femur weights were greater (P < 0.05) for T3, T4, and T5 compared to T2, which had the lowest tibia ash content (P < 0.05) and femur geometric properties (P < 0.001). Greater antibodies to infectious bronchitis virus (P < 0.01) and Ca retention (P < 0.001) were observed for T3 and T4 in comparison to T2. Based on the findings of this research, CeltiCal can adequately replace a considerable portion of limestone in broiler reduced-Ca diets containing PHY.


S-phenylmercapturic acid (S-PMA) levels in urine as an indicator of exposure to benzene in the Kinshasa population.
Data on human exposure to chemicals in Africa are scarce. A biomonitoring study was conducted in a representative sample of the population in Kinshasa (Democratic Republic of Congo) to document exposure to benzene. S-phenylmercapturic acid (S-PMA) was measured by LC-MS/MS in spot urine samples from 220 individuals (50.5% women), aged 6-70 years living in the urban area and from 50 additional subjects from the sub-rural area of Kinshasa. Data were compiled as arithmetic means, geometric means, percentile 95th and range expressed in μg/L. Overall, living in urban Kinshasa was associated with increased levels of S-PMA in urine as compared to a population living in the sub-rural area. Increased levels were also found by comparison with some date from literature. This study reveals the high benzene exposure of the Kinshasa population requiring the determination of benzene concentrations in ambient air of Kinshasa and limit values for the protection of human health.


Generation of Avian Induced Pluripotent Stem Cells.
Avian species are among the most diverse vertebrates on our planet and significantly contribute to the balance of the ecology. They are also important food source and serve as a central animal model to decipher developmental biology and disease principles. Derivation of induced pluripotent stem cells (iPSCs) from avian species would enable conservation of genetic diversity as well as offer a valuable cell source that facilitates the use of avian models in many areas of basic and applied research. In this chapter, we describe methods used to successfully reprogram quail fibroblasts into iPSCs by using human transcription factors and the techniques critical to the characterization of their pluripotency.


Morphometric evaluation of rhesus articular cartilage: correlation of hexosamine and 35SO4 = incorporation.
Morphometric evaluation of 20 rhesus articular cartilage samples were correlated with hexosamine and an 18-hour incorporation of 35SO4- as a measure of proteoglycan production on samples from the same joint. Reduced chondrocyte cellularity was the basis of the reduced maintenance of the matrix by the chondrocyte. In the more cellular cartilage, the matrix/lacunae area ratio was less than 25, and the less cellular group had a ratio of greater than 40. An inverse correlation existed between morphometric cartilage matrix/lacunar area ratio and hexosamine content. A significant difference of 35SO4 incorporation was not seen between the three morphometric grades. Morphometric assessment reduces the subjectivity of articular cartilage evaluation.


An alternative comprehensive index to quantify the interactive effect of temperature and relative humidity on hand, foot and mouth disease: A two-stage time series study including 143 cities in mainland China.
Comprehensive indices have been used to quantify the interactive effect of temperature and humidity on hand, foot and mouth disease (HFMD). The majority of them reflect how weather feels to humans. In this study, we propose an alternative index aiming to reflect the impacts of weather on HFMD and compare its performance with that of previous indices. We proposed an index defined as the product of temperature and a weight parameter raised to the rescaled relative humidity, denoted by THIa. We then compared its model fit and heterogeneity with those of previous indices (including the humidex, heat index and temperature) by a multicity two-stage time series analysis. We first built a common distributed lag nonlinear model to estimate the associations between different indices and HFMD for each city separately. We then pooled the city-specific estimates and compared the average model fit (measured by the QAIC) and heterogeneity (measured by I2) among the different indices. We included the time series of HFMD and meteorological variables from 143 cities in mainland China from 2009 to 2014. By varying the weight parameter of THIa, the results suggested that 100% relative humidity can amplify the effects of temperature on HFMD 1.6-fold compared to 50% relative humidity. By comparing different candidate indices, THIa performed the best in terms of the average of the model fits (QAIC = 9449.37), followed by humidex, heat index and temperature. In addition, the estimated exposure-response curves between THIa and HFMD were consistent across climate regions with minimum heterogeneity (I2 = 65.90), whereas the others varied across climate regions. This study proposed an alternative comprehensive index to characterize the interactive effects of temperature and humidity on HFMD. In addition, the results also imply that previous human-based indices might not be sufficient to reflect the complicated associations between weather and HFMD.


Development of a stickleback kidney cell culture assay for the screening of androgenic and anti-androgenic endocrine disrupters.
Issues raised by the presence in the environment of chemicals able to mimic or antagonize the action of androgenic hormones are of growing concern. Here we report the development of a novel in vitro test for the screening of (anti-)androgenic chemicals, based on primary cultures of stickleback kidney cells that produce a protein, the spiggin, in response to androgenic stimulation. Cell spiggin content was measured by ELISA. Comparison between cell cultures from quiescent males, photoperiodically stimulated males, control females and dihydrotestosterone (DHT)-primed females led to the selection of cell cultures from DHT-primed females for the development of a standardized protocol. 48h of treatment with androgens proved to be sufficient to induce concentration-dependent increase in spiggin cell content with a high sensitivity. DHT induced a significant spiggin increase at 10(-12)M, while testosterone (T) and the teleost specific androgen 11-ketotestosterone (11-KT) had a significant effect at 10(-10)M. Maximal responses were obtained with 10(-8)M DHT and 10(-6)M T and 11-KT. This indicates a higher sensitivity to DHT than to T and 11-KT, in agreement with previous data on stickleback kidney androgen receptor affinity. No effect was observed with other steroids or thyroid hormone, indicating the androgen specificity of the test. The anabolic steroid 17beta-Trenbolone (TB) was able to stimulate spiggin synthesis in a concentration-dependent manner with a significant effect at a concentration as low as 10(-10)M, and a maximal effect at 10(-6)M. The synthetic human androgen receptor antagonist, flutamide had no effect alone, but concentration-dependently inhibited the stimulatory effect of 10(-8)M 11-KT with a complete inhibition at 10(-6)M flutamide. This cell culture system provides an innovative tool for the rapid and sensitive screening of androgenic and anti-androgenic properties of environmental contaminants.


Improved methodology for detecting aflatoxin production quantitatively in natural media.
This report describes a simple, rapid, and quantitative method for screening the aflatoxin production by moulds of theAspergillus flavus group, using a natural media (moist wheat or rice), and a single chloroform extraction for aflatoxins. The aflatoxins were detected and quantified by thin layer chromatography. The methodology proposed was useful in detecting aflatoxin production on the 3rd day of incubation in more than 85 % of the strains studied (both culture collection strains and field isolates).


4-Hydroxybenzoic acid (4-HBA) enhances the sensitivity of human breast cancer cells to adriamycin as a specific HDAC6 inhibitor by promoting HIPK2/p53 pathway.
Breast cancer is reported a very complex disease along with heterogeneous morphological characteristics and unrelated clinical behavior, and is a leading cancer among female. Nevertheless, chemo-resistance is frequently observed. Adriamycin (ADM) is a always employed drug to treat clinical breast cancer. However, strong resistance to ADM limited its clinical efficacy. Deregulation of HDAC6 activity is linked to various diseases including cancer resulting in accumulating interest for developing HDAC6 inhibitors. In the present study, for the first time, we found that 4-Hydroxybenzoic acid (4-HBA), as histone deacetylase 6 (HDAC6) inhibitor, could successfully reverse ADM resistance in human breast cancer cells. 4-HBA significantly promoted the anticancer effect of ADM on apoptosis induction, as evidenced by the increased expressions of Caspase-3 and PARP cleavage, which was associated with the promotion p53 and homeodomain interacting protein kinase-2 (HIPK2) expressions in ADM-resistant breast cancer cells. Furthermore, the suppressive effect of ADM on drug-resistant breast cancer cells was accelerated by 4-HBA through increasing the number of cells distributed in G2/M phase of cell cycle arrest. Inhibiting HIPK2/p53 pathway could abolish 4-HBA/ADM co-treatment-induced apoptosis and G2/M cell cycle arrest. Importantly, HDAC6 expressions were also significantly down-regulated in ADM-resistance breast cancer cells co-treated with ADM and 4-HBA. Additionally, 4-HBA clearly potentiated the anticancer role of ADM in the MCF-7 breast cancer animal model with low toxicity. Therefore, 4-HBA could be applied as an effective HDAC6 inhibitor to reverse human breast cancer resistance. Herein, the 4-HBA and ADM combination might represent as a useful therapeutic strategy to prevent human breast cancer.


Novel nano-insulin formulation modulates cytokine secretion and remodeling to accelerate diabetic wound healing.
Little is known about insulin's wound healing capability in normal as well as diabetic conditions. We here report specific interaction of silver nanoparticles (AgNPs) with insulin by making a ~2 nm thick coat around the AgNPs and its potent wound healing efficacy. Characterization of the interaction of human insulin with silver nanoparticles showed confirmed alteration of amide-I in insulin whereas amide-II and III remained unaltered. Further, nanoparticles protein interaction kinetics showed spontaneous interaction at physiological temperature with ΔG, ΔS, Ea and Ka values -7.48, 0.076, 3.84 kcal mol-1 and 6 × 105 s-1 respectively. Insulin loaded AgNPs (IAgNPs) showed significant improvement in healing activity in vitro (HEKa cells) and in vivo (Wister Rats) in comparison with the control in both normal and diabetic conditions. The underlying mechanism was attributed to a regulation of the balance between pro (IL-6, TNFα) and anti-inflammatory cytokines (IL-10) at the wound site to promote faster wound remodeling.


Mannitol upregulates monocyte HLA-DR, monocyte and neutrophil CD11b, and inhibits neutrophil apoptosis.
D-Mannitol is a substance widely used for both clinical applications as a strong diuretic and in basic research as a purportedly inert osmotic control substance. However, recent experiments have shown that mannitol is able to decrease neutrophil apoptosis in vitro by more than 25%. The aim of the current study was to assess mannitol's effects on two immune cell activation markers; CD11b on neutrophils and monocytes, and HLA-DR on monocytes. Exposure of diluted whole blood (1:10 in RPMI 1640) to increasing concentrations of mannitol for 24 h was associated with a significant increase in both monocyte and neutrophil CD11b expression in non-lipopolysaccharide (LPS)-stimulated samples. Monocyte HLA-DR, but not neutrophil HLA-DR, was significantly higher in the presence of 16.5 mM/l mannitol, whereas isoosmolar NaCl had no effect. Levels of IL-6 and TNF-alpha were not affected by mannitol in our model. All reagents were tested negative for endotoxin contamination. Together, these data indicate that mannitol may directly interact with neutrophils and monocytes. Further systematic evaluation is indicated to define the precise immunomodulating actions of mannitol, and to assess these effects in patients receiving such therapy.


Regional specialization in the Malpighian tubules of the New Zealand glow-worm Arachnocampa luminosa (Diptera: mycetophilidae). The structure and function of type I and II cells.
The Malpighian tubules of the glow-worm Arachnocampa luminosa are divided into four morphologically distinct regions (Parts 1--4) each comprised of a different cell type (Types I--IV). The ultrastructure of Type II cells is indicative of a transport function. The basal cell surface is highly invaginated and at the apical surface the lumen is lined with microvilli about 80% of which contain mitochondria. Spherites contained in these cells are formed from small vesicles produced by the Golgi apparatus. They have a central uric acid core enclosed by laminations of phosphates of calcium and magnesium. Cells of Part 2 of the tubule secrete a fluid high in potassium (173 mM) and low in sodium (18 mM). The cell is 30 mV negative and the lumen 44 mV positive to the bathing solution. This is consistent with the proposal of an apical cation pump. The secretion produced by Part 2 of the tubules is modified by the Type I cells by the reabsorption of potassium (162 mM) and the addition of sodium (24 mM) to the primary excretory fluid. Type I cells are 20 mV negative and the lumen 22 mV positive with respect to the bathing medium. From ultrastructural observations, Type I cells exhibit features characteristic of transporting cells thought to have an absorptive function. The basal and apical cell surfaces are extensively folded, and mitochondria are found in bands above the basal infoldings and below the microvilli. Mitochondria do not penetrate the microvilli. On comparative grounds, the fine structure of Type I cells suggest that they reabsorb ions from the tubule lumen. Energy for these processes may come from the breakdown of lipids by microperoxisomes contained within these cells. Alternatively, the fluid produced by Part 2 of the tubule may be modified passively by diffusional processes across Type I cells.


Interactions of translational factor EF-G with the bacterial ribosome before and after mRNA translocation.
A conserved translation factor, known as EF-G in bacteria, promotes the translocation of tRNA and mRNA in the ribosome during protein synthesis. Here, EF-G.ribosome complexes in two intermediate states, before and after mRNA translocation, have been probed with hydroxyl radicals generated from free Fe(II)-EDTA. Before mRNA translocation and GTP hydrolysis, EF-G protected a limited set of nucleotides in both subunits of the ribosome from cleavage by hydroxyl radicals. In this state, an extensive set of nucleotides, in the platform and head domains of the 30S subunit and in the L7/L12 stalk region of the 50S subunit, became more exposed to hydroxyl radical attack, suggestive of conformational changes in these domains. Following mRNA translocation, EF-G protected a larger set of nucleotides (23S rRNA helices H43, H44, H89, and H95; 16S rRNA helices h5 and h15). No nucleotide with enhanced reactivity to hydroxyl radicals was detected in this latter state. Both before and after mRNA translocation, EF-G protected identical nucleotides in h5 and h15 of the 30S subunit. These results suggest that h5 and h15 may remain associated with EF-G during the dynamic course of the translocation mechanism. Nucleotides in H43 and H44 of the 50S subunit were protected only after translocation and GTP hydrolysis, suggesting that these helices interact dynamically with EF-G. The effects in H95 suggest that EF-G interacts weakly with H95 before mRNA translocation and strongly and more extensively with this helix following mRNA translocation.


Lung cysts in subacute hypersensitivity pneumonitis.
To determine the frequency of cystic lesions on high-resolution computed tomography (CT) in patients with subacute hypersensitivity pneumonitis. High-resolution CT scans in 182 patients with proven subacute hypersensitivity pneumonitis were retrospectively evaluated for the presence of lung cysts. Patients with CT evidence of emphysema or interstitial fibrosis and patients with connective tissue disease were excluded from the study. Two thoracic radiologists reviewed the thoracic CT scans for the presence and location of lung cysts. Thin-walled lung cysts were identified in 24 (13%) of 182 patients and ranged in size from 3 to 25 mm in maximal diameter. The patients included 16 men and 8 women (age range, 30-79 years; mean age, 48 years). The cysts ranged from 1 to 15 in number (mean, 4 cysts) and had a random distribution. Sixteen of the 24 patients also had areas of decreased attenuation and vascularity consistent with air trapping. Thin-walled cysts can be seen in a small percentage of patients with subacute hypersensitivity pneumonitis. The cysts resemble those seen in lymphocytic interstitial pneumonia, and their pathogenesis is uncertain.


Angiotensin-converting enzyme insertion/deletion polymorphism is a risk factor for thoracic aortic aneurysm in patients with bicuspid or tricuspid aortic valves.
The angiotensin-converting enzyme (ACE) is highly expressed in the aneurysmal vascular wall, in both animal models and human disease. Genetic variations in ACE could be crucial in determining the risk of thoracic aortic aneurysm (TAA). The aim of the present study was to examine the role of ACE insertion/deletion polymorphism on the risk of TAA in patients with bicuspid aortic valves or tricuspid aortic valves. We enrolled 216 patients (158 men; age, 58.9±14.9 years) with TAA, associated with bicuspid aortic valves (n=105) and tricuspid aortic valves (n=111) compared with 312 patients (252 men; age, 54.6±11.0 years) with angiographically proven coronary artery disease and 300 healthy controls (91 men; age, 40.4±10.5 years). The genotype distribution of ACE insertion/deletion was significantly different between the patients with TAA compared with both the control group (P=.0005) and the coronary artery disease group (P=.03). The genotypes were not different between the control group and the coronary artery disease group (P=.3). Compared with the controls, both the bicuspid aortic valve patients (P=.0008) and tricuspid aortic valve patients (P<.0001) had a greater frequency of allele D. The aortic diameters were significantly different among the three genotypes (48.3±6.6, 45.3±8.9, 39.9±8.7 for the DD, DI, and II genotypes, respectively; P=.0002). A synergistic effect between the ACE D allele and hypertension was found for both an increased aortic diameter (P=.003) and the risk of TAA (P<.001). On multivariate logistic regression analysis, D allele (odds ratio, 3.0; 95% confidence interval, 1.1-8.1; P=.03) was a significant predictor of TAA. ACE insertion/deletion polymorphism represents a genetic biomarker for TAA. These findings could have a significant effect on both the early detection and effective pharmacologic treatment of aortic disease.


EEG-LORETA endophenotypes of the common idiopathic generalized epilepsy syndromes.
We tested the hypothesis that the cortical areas with abnormal local EEG synchronization are dissimilar in the three common idiopathic generalized epilepsy (IGE) phenotypes: IGE patients with absence seizures (ABS), juvenile myoclonic epilepsy (JME) and epilepsy with generalized tonic-clonic seizures exclusively (EGTCS). Groups of unmedicated ABS, JME and EGTCS patients were investigated. Waking EEG background activity (without any epileptiform potentials) was analyzed by a source localization method, LORETA (Low Resolution Electromagnetic Tomography). Each patient group was compared to a separate, age-matched group of healthy control persons. Voxel-based, normalized broad-band (delta, theta, alpha, and beta) and very narrow band (VNB, 1Hz bandwidth, from 1 to 25Hz) LORETA activity (=current source density, A/m(2)) were computed for each person. Group comparison included subtraction (average patient data minus average control data) and group statistics (multiple t-tests, where Bonferroni-corrected p<0.05 values were accepted as statistically significant). Statistically not significant main findings were: overall increased delta and theta broad band activity in the ABS and JME groups; decrease of alpha and beta activity in the EGTCS group. Statistically significant main findings were as follows. JME group: bilaterally increased theta activity in posterior (temporal, parietal, and occipital) cortical areas; bilaterally increased activity in the medial and basal prefrontal area in the 8Hz VNB; bilaterally decreased activity in the precuneus, posterior cingulate and superior parietal lobule in the 11Hz and 21-22Hz VNBs. ABS group: bilaterally increased theta activity emerged in the basal prefrontal and medial temporal limbic areas. Decreased activity was found at 19-21Hz in the right postcentral gyrus and parts of the right superior and medial temporal gyri. EGTCS group: decreased activity was found in the frontal cortex and the postcentral gyrus at 10-11Hz, increased activity in the right parahippocampal gyrus at 16-18Hz. Increased theta activity in the posterior parts of the cortex is the endophenotype for JME. Increased theta activity in the fronto-temporal limbic areas is the endophenotype for ABS. Statistically not significant findings might indicate diffuse biochemical abnormality of the cortex in JME and ABS. EEG-LORETA endophenotypes may correspond to the selective propensity to generate absence and myoclonic seizures in the ABS and JME syndromes.


Prognostic value of cardiopulmonary exercise testing repetition during follow-up of clinically stable patients with severe dilated cardiomyopathy. A preliminary study.
Cardiopulmonary exercise testing (CPET) is a recognized tool for prognostic stratification in patients with dilated cardiomyopathy (DCM). Given the lack of data currently available, the aim of this study was to test the prognostic value of repeating CPET during the follow-up of patients with DCM. This multicenter, retrospective study, analyzed DCM patients who consecutively performed two echocardiographies and CPETs during clinical stability. The study end-point was a composite of death from all causes, heart transplantation, left ventricular assist device implantation, life-threatening ventricular arrhythmias or hospitalization for heart failure. 216 DCM patients were enrolled (52 years, 78% male, NYHA I-II 82%, LVEF 32%, 94% on ACE inhibitors/ARNI, 95% on beta-blockers). The interval between CPETs was 15 months. During a median follow-up of 38 months from the second CPET, 102 (47%) patients experienced the study end-point. Among them, there was stability of echocardiographic values but a significant worsening of functional capacity. Among the 173 patients (80%) who did not show echocardiographic left ventricular reverse remodeling (LVRR), the 1-year prevalence of the study-end point was higher in patients who worsened vs patients who maintained stable their functional capacity at CPET (38 vs. 15% respectively, p-value: 0.001). These results were consistent also when excluding life-threatening ventricular arrhythmias from the composite end-point. In clinically stable DCM patients with important depression of LVEF, the repetition of combined echocardiography and CPET might be recommended. When LVRR fails, 1-year repetition of CPET could identify higher-risk patients.


Deficient T(H)-1 responses from TNF-alpha-matured and alpha-CD40-matured dendritic cells.
The ultimate success of dendritic cell (DC) vaccination for the active immunotherapy of neoplasia is thought to be dependent on a very large number of variables, including DC generation protocol, loading methodology, dose, route of administration, and maturation method. Although the use of a maturation cocktail comprising interleukin (IL)-1beta, tumor necrosis factor-alpha, IL-6, and prostaglandin E2 (ITIP) has recently appeared in the literature, much of the data in the basic and clinical literature have been generated using DCs matured with the single inflammatory cytokine TNF-alpha. Here, we demonstrate that DCs matured with TNF-alpha alone or in combination with CD40 agonism are highly deficient, both physiologically and functionally, in comparison with DCs matured with IL-1beta, TNF-alpha, IL-6, and prostaglandin E2. Empirically, the data suggest that DCs matured with these agents are deficient in the induction of type 1 T-helper responses. We further speculate that DCs matured by these methods might be suboptimal for the priming of naive responses.


Osteoporosis, calcium requirement, and factors causing calcium loss.
A large percentage of subjects receiving a normal calcium intake of 800 mg per day was in negative calcium balance and increasing the calcium intake to various levels up to 2200 mg per day showed a plateau of the calcium balance at the 1200 mg per day intake, indicating a threshold of calcium absorption at this intake level. Dietary factors, such as a phosphorus intake up to 2000 mg per day and complex proteins, such as red meat, do not cause calcium loss. Several commonly used drugs induce significant calcium loss, particularly aluminum-containing antacids, and, if used for prolonged periods of time, can contribute to the development of osteoporosis.


Safety and peri-operative outcomes during learning curve of robot-assisted laparoscopic prostatectomy: a multi-institutional study of fellowship-trained robotic surgeons versus experienced open radical prostatectomy surgeons incorporating robot-assisted laparoscopic prostatectomy.
To analyze and compare the safety and peri-operative outcomes of fellowship-trained robotic surgeons (FEL) and experienced open surgeons (OE) incorporating robot-assisted laparoscopic prostatectomy (RALP) into practice. Multiinstitutional, prospective data were collected on the first 30 RALP performed by FEL and OE (defined as over 1000 prostatectomies) incorporating RALP into practice. Morbidity from the peri-operative course was evaluated as were operative outcomes. The second 30 cases from the OE group were evaluated to assess for improvement with experience. There were no rectal injuries or death in either group. Blood transfusion rates did not differ between the two groups (2% vs. 3%, p = 0.65). Open conversion occurred three times in the OE group but only within the first 30 cases. In the first 30 cases FEL had statistically lower rates of positive margins (15% vs. 34%, p = 0.008) and decreased likelihood of prolonged urethral catheter leakage (5% vs. 19%, p = 0.009). The FEL group had lower rates of failure of prostate-specific antigen to nadir < 0.15 ng/mL (2% vs. 10%, p = 0.056). There were no reoperations in the FEL group but present in 2% of the OE group initially. The second 30 cases of the OE group noted a statistical improvement for all parameters with margin rates and the requirement of prolonged catheterization becoming statistically comparable to those of the FEL group. OE can safely incorporate RALP into practice and achieve outcomes comparable to FEL quickly. As anticipated, FEL achieve these endpoints earlier in their practice.


Effect of tobacco smoking on various histological types of lung cancer.
In a population-based case/control study the differential lung cancer risk patterns due to tobacco smoking habits of various histological types have been investigated. The cases were 1432 deaths from lung cancer in the years 1980-1987, of which the histological type was known for 627 individuals. There was 54% squamous cell carcinoma, 24% small-cell carcinoma and 17% adenocarcinoma. Controls were 1343 deaths from other causes. Next-of-kin interviews were performed. The results of the study confirmed that cigarette smoking is associated with all histological types of lung cancer; however, the dose/response relationship between smoking and adenocarcinoma differed clearly from that observed in squamous and small-cell carcinomas. In the latter histological types the gradient of risk was much stronger as the number of cigarettes smoked or duration of smoking increased. The overall relative risk for smoking in small-cell and squamous cell carcinoma was 15.4 and 13.5 respectively, whereas that for adenocarcinoma was weaker (relative risk = 3.1). An interesting difference between squamous and small-cell carcinomas was found also for patients who gave up smoking. The effect of stopping was more pronounced in squamous cell carcinoma. The attributable risks for smoking in squamous and small-cell carcinoma were much higher (90% and 88% respectively) than for adenocarcinoma (64%). The data suggest that adenocarcinoma is likely to be related to other factors than tobacco smoking to a greater extent than are squamous or small cell carcinoma. Possible sources of bias, such as missing histological diagnoses, are discussed in detail.


Chemiluminescence of doped carbon dots with H2O2-KMnO4 system for the detection of Cu2+ and tannin.
The carbon dots doped with chlorine and phosphorus (CDs-Cl,P) were used as chemiluminescence (CL) reagent for the sensitive detection of copper ions (Cu2+) and tannin (TA). The CDs-Cl,P was found to strongly enhance the reaction of H2O2 and KMnO4 in alkaline medium. The enhanced CL behavior of CDs-Cl,P was investigated and it was found that some radicals such as •OH, •O2- and 1O2 appeared in the CL reaction process. The participation of Cu2+ could result in an enhanced CL intensity of the CDs-Cl,P-H2O2-KMnO4 system due to the Cu2+-catalyzed decomposition of H2O2 resulting in more •OH generation. Therefore, the CDs-Cl,P-H2O2-KMnO4 system was used to selectively quantify Cu2+ in solution by CL emission. A linear increase was observed between CL intensity and Cu2+ concentration. The CDs-Cl,P-H2O2-KMnO4 system allowed the detection of Cu2+ down to lower concentration of 0.1 μM with a linear range of 0.2-60.0 μM. Moreover, TA as a common polyphenolic compound, could selectively decrease the CL signal of the CDs-Cl,P-H2O2-KMnO4-Cu2+ system due to its complexation with Cu2+. On this basis, the CL assay for TA was also developed. The detection limit was 0.14 μM and the linear range was from 5.0 μM to 100.0 μM. The proposed method was successfully applied to the determination of Cu2+ and TA in water, rice dumplings leaves, sodium copper chlorophyllin and wine samples with satisfactory results.


Antiproliferative mechanism of anti-class II monoclonal antibodies.
The function of Class II molecules in proliferation was explored by treating human cell lines with three distinct anti-DR monoclonal antibodies (MABs). Dose-dependent, specific inhibition of eight DR+ cell lines of different origin and lineage was found. Inhibition was durable (i.e., cells did not become resistant to the anti-DR MABs despite prolonged treatment) yet reversible. The mechanism of inhibition was not due to differentiation or killing but was cytostatic. Inhibition was temporally associated with decreases in nuclear size and irregularity and appeared to be due to a non-phase-specific cell cycle arrest.


Bimetallic NiPt nanoparticles-enhanced catalyst supported on alginate-based biohydrogels for sustainable hydrogen production.
Alginate hydrogel beads were loaded with bimetallic NiPt nanoparticles by in situ reduction of the respective polymer matrix containing precursor metallic ions using a NaBH4 aqueous solution. The alginate hydrogel beads loaded with NiPt nanoparticles were characterized by TEM, AAS, FT-IR, TGA, XPS, and oscillatory rheometry. The prepared hybrid hydrogels were proven to be effective as catalytic materials for the hydrolysis of ammonia borane (AB) for quantitative hydrogen generation using catalytic loadings of 0.1 mol%. In addition, the reaction mechanism of the hydrolytic reaction using NiPt loaded alginate hydrogel beads was determined by Langmuir-Hinshelwood model. The experimental results showed that the reaction mechanism consisted of an initial fast adsorption of reactants at the surface of the nanoparticles, followed by a rate-limiting surface reaction. The NiPt nanoalloys exhibited an enhanced behavior for hydrogen generation with a maximum TOF of 84.1 min-1, almost 71 % higher compared to monometallic platinum atoms, and likely related to a synergistic interaction between both metals. Finally, the hydrogel matrix enabled the material to be easily recovered from the reaction medium and reused in further catalytic cycles without desorption of active nanoparticles from the material.


Spinal Cord Injury-Functional Index/Capacity: Responsiveness to Change Over Time.
To establish responsiveness of 3 Spinal Cord Injury-Functional Index/Capacity (SCI-FI/C) item banks in the first year after spinal cord injury (SCI). Longitudinal patient-reported outcomes assessment replicated through secondary analysis of an independent data set. A total of 8 SCI Model Systems rehabilitation hospitals in the United States. Study 1 participants included 184 adults with recent (≤4 months) traumatic SCI and 221 community-dwelling adults (>1 year post injury) (N=405). Study 2 participants were 418 individuals with recent SCI (≤4 months) (N=418). In study 1, SCI-FI/C computer adaptive tests were presented in a standardized interview format either in person or by phone call at baseline and 6-month follow-up. Responsiveness was examined by comparing 6-month changes in SCI-FI scores within and across samples (recently injured vs community-dwelling) because only the recent injury sample was expected to exhibit change over time. Effect sizes were also computed. In study 2, the study 1 results were cross-validated in a second sample with recent SCI 1 year after baseline measurement. Study 2 also compared the SCI-FI/C measures' responsiveness to that of the Self-reported Functional Measure (SRFM) and stratified results by injury diagnosis and completeness. The SCI-FI Basic Mobility/C, Self-care/C and Fine Motor/C item banks (study 1 and study 2); Self-reported Functional Measure SRFM (study 2 only). In study 1, changes in SCI-FI/C scores between baseline and 6-month follow-up were statistically significant (P<.01) for recently injured individuals. SCI-FI Basic Mobility/C, Self-care/C, and Fine Motor/C item banks demonstrated small to medium effect sizes in the recently injured sample. In the community-dwelling sample, all SCI-FI/C effects were negligible (ie, effect size<0.08). Study 2 results were similar to study 1. As expected, SCI-FI Basic Mobility/C and Self-care/C were responsive to change for all individuals in study 2, whereas the SCI-FI Fine Motor/C was responsive only for individuals with tetraplegia and incomplete paraplegia. The SRFM demonstrated a medium effect size for responsiveness (effect size=0.65). The SCI-FI Basic Mobility/C and Self-care/C banks demonstrate adequate sensitivity to change at 6 months and 1 year for all individuals with SCI, while the SCI-FI/C Fine Motor item bank is sensitive to change in individuals with tetraplegia or incomplete paraplegia. All SCI-FI/C banks demonstrate stability in a sample not expected to change. Results provide support for the use of these measures for research or clinical use.


In-hospital monitoring of T-wave alternans in a case of amiodarone-induced torsade de pointes: clinical and methodologic insights.
We report a case of macroscopic T-wave alternans occurring 30 min before the onset of amiodarone-induced torsade de pointes, illustrating a means to monitor for proarrhythmia.


Improved Androgen Receptor Splice Variant 7 Detection Using a Highly Sensitive Assay to Predict Resistance to Abiraterone or Enzalutamide in Metastatic Prostate Cancer Patients.
In metastatic castration-resistant prostate cancer (mCRPC), androgen receptor splice variant 7 (AR-V7) expression is associated with a low response to androgen receptor signaling (ARS) inhibitors such as abiraterone or enzalutamide. To perform a highly sensitive assay for detecting AR-V7 (hsAR-V7) in circulating tumor cells (CTCs) and evaluate its ability to predict response to ARS inhibitors. From 41 mCRPC patients, CTCs were prospectively enriched using AdnaTest platform and analyzed for AR-V7 with and without the highly sensitive assay. The first objective of the study was to compare AR-V7 detection rates with and without the highly sensitive assay. Next, we investigated how AR-V7 (detected without the highly sensitive assay) and hsAR-V7 status influenced prostate-specific antigen (PSA) response and long-term clinical outcomes (PSA progression-free survival [PFS] and radiological PFS) after ARS-inhibitor treatment. Finally, discriminatory abilities of the assays were assessed by C-index to compare their impact on long-term clinical outcomes. AR-V7 detection rates increased from 22% to 56% when the highly sensitive assay was used. The discriminatory abilities of hsAR-V7 for PSA PFS (C-index, 0.74; 95% confidence interval [CI], 0.60-0.88) and radiological PFS (0.70; 95% CI, 0.55-0.85) were higher than those of AR-V7 detected without the highly sensitive assay (0.60, 0.51-0.72, and 0.56, 0.44-0.67, respectively). After ARS-inhibitor treatment, PSA response was lower in hsAR-V7+ (53%) than in hsAR-V7- (93%) patients (p = 0.016). AR-V7+ patients had shorter median PSA PFS (3.0 vs 10.6 mo, p = 0.032) and nonsignificantly shorter median radiological PFS (6.0 vs 14.8 mo, p = 0.24) compared with AR-V7- patients. The hsAR-V7+ status was associated with shorter median PSA PFS (3.0 mo vs not reached, p = 0.0001) and radiological PFS (median, 6.0 mo vs not reached, p = 0.0026). The hsAR-V7 assay achieved the highest AR-V7 detection rates among those reported in mCRPC. Discriminatory abilities for long-term clinical outcomes were better with hsAR-V7 assay. We prospectively analyzed circulating tumor cells from men with metastatic castration-resistant prostate cancer for androgen receptor splice variant 7 (AR-V7) status using a highly sensitive assay. It yielded higher AR-V7 detection rates and predicted resistance to androgen receptor signaling inhibitors with better discriminatory abilities for long-term clinical outcomes.


Effect of microhabitat on fitness components of larval tiger salamanders, Ambystoma tigrinum nebulosum.
Survivorship and growth of larval tiger salamanders, Ambystoma tigrinum nebulosum, in the White Mountains of east-central Arizona were compared in six microhabitats using field enclosures during summers, 1983-85. Microhabitats were vegetated and nonvegetated shallows, surface, middle, and bottom horizontal layers of limnetic areas, and the vertical limnetic water column. Initial enclosure densities (0.025 larvae per 1) were identical among microhabitats. Three enclosures were placed in each microhabitat in two ponds. Larval survivorship and growth were usually greatest in vegetated shallows in lowest in middle and bottom limnetic enclosures, despite several population dieoffs. Lower fitness, as reflected in survivorship and growth, in these latter enclosures was correlated with lower food levels, temperatures, and oxygen concentrations in deeper limnetic areas. Relative fitness varied greatly between years while food levels, temperatures, and oxygen concentrations within microhabitats remained relatively constant indicating additional factors affected fitness. Disparities in fitness between microhabitats apparently affect habitat use patterns of larvae.


How can the potential of the duocarmycins be unlocked for cancer therapy?
The duocarmycins belong to a class of agent that has fascinated scientists for over four decades. Their exquisite potency, unique mechanism of action, and efficacy in multidrug-resistant tumour models makes them attractive to medicinal chemists and drug hunters. However, despite great advances in fine-tuning biological activity through structure-activity relationship studies (SARS), no duocarmycin-based therapeutic has reached clinical approval. In this review, we provide an overview of the most promising strategies currently used and include both tumour-targeted prodrug approaches and antibody-directed technologies.


TERT mutations correlate with higher TMB value and unique tumor microenvironment and may be a potential biomarker for anti-CTLA4 treatment.
Immune checkpoint inhibitors (ICIs) have recently changed therapeutic paradigms for patients across multiple cancer types. However, current biomarkers cannot accurately predict responses to ICIs. Telomerase reverse transcriptase (TERT) mutations lead to an aberrant upregulation of TERT expression, and ultimately allow telomere maintenance, thus supporting immortalization of cancer cells. This study aimed to investigate whether the TERT mutation is a potential predictor of ICI treatment across all cancer types. TERT mutations positively correlated with a higher tumor mutational burden (TMB) value, neoantigen load, and tumor purity. Lymphocyte infiltration, macrophage regulation, interferon-gamma (IFN-γ) response, and transforming growth factor-β (TGF-β) response which was representative immune-expression signatures, all had higher signature scores in the TERT mutation group. Activated CD4 T cell, naïve B cell, activated dendritic cell, M0 macrophage, M1 macrophage, neutrophil, resting NK cell, and plasma cells all had relatively higher immune scores in the TERT mutation group, whereas Th series cells, memory B cell, resting mast cells, monocytes, and activated NK cells had lower immune scores. Notably, in the subgroup analysis of monotherapy and combination ICI treatment, only in the anti-cytotoxic-T-lymphocyte-associated antigen 4 (anti-CTLA4) group, patients with TERT mutations had a better prognosis, especially for melanoma. Therefore, TERT mutations were closely related to a higher TMB value and unique tumor microenvironment, which may be the reason that TERT mutations may be a potential biomarker for anti-CTLA4 treatment.


Practice nurses' communication with people living with type 2 diabetes: A scoping review.
To identify the key characteristics of practice nurses' communication with people living with Type 2 Diabetes (T2D) where lifestyle activities are discussed. A scoping review of the peer-reviewed literature was conducted. Reflexive thematic analysis was used to identify key themes that emerged. The PRISMA-ScR checklist was followed. 25 studies were retained in the final review. Nurses who were committed to engaging in lifestyle discussions created supportive environments and built rapport to enable change conversations. However, this was present in just 20% of the studies. In most studies, (60%) nurses continued to use traditional health education communication styles, had little understanding of behaviour change theories, lacked skills in behaviour change counselling and were reluctant to engage in behaviour change discussions with people with T2D . Nurses require a deeper understanding of behavioural change theories and skills in behavioural counselling. Practice nurses have a unique opportunity to facilitate T2D remission by engaging in evidence-based behaviour change communication. A behaviour change training intervention is needed that recognises the environment of practice nurse consultations. It needs to be pragmatic and fully consider the enablers and barriers to addressing behaviour change in both the nurse and the person with T2D.


Corrigendum.
Corrigendum to "Rationale and design of the NAGOYA HEART Study: Comparison between valsartan and amlodipine regarding morbidity and mortality in patients with hypertension and glucose intolerance" [J. Cardiol. 56 (2010) 111–117] The authors of the manuscript would like to correct affiliation of Mr. Nobuo Shirahashi, who was mentioned in the Appendix section of this article. The correct affiliation of Mr. Shirahashi was Novartis Pharma KK and Department of Preventive Medicine and Environmental Health, Osaka City University Medical School, Osaka, Japan.


Back Pain: Differential Diagnosis and Management.
Back pain is a common condition affecting millions of individuals each year. A biopsychosocial approach to back pain provides the best clinical framework. A detailed history and physical examination with a thorough workup are required to exclude emergent or nonoperative etiologies of back pain. The treatment of back pain first uses conventional therapies including lifestyle modifications, nonsteroidal anti-inflammatory drugs, physical therapy, and cognitive behavioral therapy. If these options have been exhausted and pain persists for greater than 6 weeks, imaging and a specialist referral may be indicated.


A comparative evaluation of the Eustachian tube cartilage between healthy and diseased ears using a 3 Tesla MRI.
The purpose of this study was to prospectively assess the Eustachian tube (ET) cartilage using 3 Tesla (3T) magnetic resonance imaging (MRI) and compare the results between healthy ears and those with a middle ear disease. The study included 56 ears with a middle ear disease as the patient group and 100 ears without a middle ear disease as the control group. The patients' age ranged from 18 to 65 years. The axial three-dimensional (3D) multiple echo recombined gradient echo (MERGE) sequence and oblique parasagittal planes were obtained. Visualisation of the ET cartilage was assessed on the MR images using a three-point numerical rating score. In the axial plane, the ET lumen's diameter was measured from the mid-portion of the cartilage. There was no significant difference between the patient group and the control group according to patients' age and gender, and the medial laminal thickness of the ET cartilage. In the patient group, the diameter of the ET cartilage was significantly smaller than in the control group. The ET lumen diameter was significantly lower according to each of the three scoring systems. 3T MRI provides an evaluation of the ET cartilage and isthmus level, which are small but important anatomical localisations and surgical landmarks. MR imaging has the potential to provide essential information on ET prior to new surgical treatments, such as balloon dilation for middle ear diseases.


Transcriptomic and genomic structural variation analyses on grape cultivars reveal new insights into the genotype-dependent responses to water stress.
Grapevine (Vitis vinifera L.) is importantly cultivated worldwide for table grape and wine production. Its cultivation requires irrigation supply, especially in arid and semiarid areas. Water deficiency can affect berry and wine quality mostly depending on the extent of plant perceived stress, which is a cultivar-specific trait. We tested the physiological and molecular responses to water deficiency of two table grape cultivars, Italia and Autumn royal, and we highlighted their different adaptation. Microarray analyses revealed that Autumn royal reacts involving only 29 genes, related to plant stress response and ABA/hormone signal transduction, to modulate the response to water deficit. Instead, cultivar Italia orchestrates a very broad response (we found 1037 differentially expressed genes) that modifies the cell wall organization, carbohydrate metabolism, response to reactive oxygen species, hormones and osmotic stress. For the first time, we integrated transcriptomic data with cultivar-specific genomics and found that ABA-perception and -signalling are key factors mediating the varietal-specific behaviour of the early response to drought. We were thus able to isolate candidate genes for the genotype-dependent response to drought. These insights will allow the identification of reliable plant stress indicators and the definition of sustainable cultivar-specific protocols for water management.


Molecular biology and genetics of gap junction channels.
Gap junctional communication between cells provides a mechanism for the movement of molecular information between cells via the unit of gap junction structure and function, the gap junctional channel. In the past five years, there has been rapid progress in identifying and characterizing a multigene family that is responsible for producing the gap junction polypeptides that are responsible for generating gap junctional channel oligomers between cells. The products of these genes have been referred to as connexins, and the multigene family can be categorized into two classes at present, the alpha class and the beta class. Members of these two classes can be distinguished on the basis of their primary sequence and overall predicted topological organization. The gap junction genes map to different chromosomes in both mice and humans, and these genes are utilized on a cell specific basis. Furthermore, these genes can be developmentally regulated, and multiple genes can be co-expressed simultaneously by the same cell type. Efforts to understand the precise structure-function relationship of the products of these different genes is now being approached by utilizing various expression systems. Criteria that can be used as a basis for determining membership in the multigene family is presented and discussed, as well as the rationale for using a nomenclature system for the gap junction multigene family that is based on genetic and structural relationships rather than the molecular size of the deduced protein products.


Non-line-of-sight snapshots and background mapping with an active corner camera.
The ability to form reconstructions beyond line-of-sight view could be transformative in a variety of fields, including search and rescue, autonomous vehicle navigation, and reconnaissance. Most existing active non-line-of-sight (NLOS) imaging methods use data collection steps in which a pulsed laser is directed at several points on a relay surface, one at a time. The prevailing approaches include raster scanning of a rectangular grid on a vertical wall opposite the volume of interest to generate a collection of confocal measurements. These and a recent method that uses a horizontal relay surface are inherently limited by the need for laser scanning. Methods that avoid laser scanning to operate in a snapshot mode are limited to treating the hidden scene of interest as one or two point targets. In this work, based on more complete optical response modeling yet still without multiple illumination positions, we demonstrate accurate reconstructions of foreground objects while also introducing the capability of mapping the stationary scenery behind moving objects. The ability to count, localize, and characterize the sizes of hidden objects, combined with mapping of the stationary hidden scene, could greatly improve indoor situational awareness in a variety of applications.


Right and left ventricular long-axis function in the fetus using angular M-mode.
Long-axis function is determined by the longitudinally oriented myocardial fibers. Postnatally, conventional M-mode is used to assess tricuspid and mitral valve ring movements in relation to the cardiac apex. During fetal life, this is precluded by variable fetal position. We assessed the feasibility of determining right and left ventricular long-axis function in the fetus. A prospective, pilot study. The four-chamber view obtained during routine fetal echocardiography was recorded in a cineloop to which B-mode guided M-mode echocardiography was applied using angular M-mode. This allowed retrospective and correct placement of the cursor line from cardiac apex to tricuspid or mitral valve rings. M-mode tracings of the valve ring movements in relation to the apex were derived from the originally stored loop. Data from 18 fetuses (17-29 weeks of gestation) were available for analysis. Total excursion of the valve rings was measured offline. A second cursor line was simultaneously placed in the left ventricular outflow tract during color flow mapping for timing purposes (n = 6). Right and left ventricular long-axis recordings were obtained in 18 and 14 cases, respectively. Total right ventricular excursion was 5.2 mm (SD, 0.9 mm) (range, 3.9-7.2 mm). Total left ventricular free wall excursion was 4.5 mm (SD, 1.1 mm) (range, 3.0-6.8 mm). For paired data, the mean of differences (right ventricle-left ventricle) was 0.8 mm (95% confidence interval 0.5-1.2). The valve rings moved towards the apex during systole (shortening) and away from it during diastole (lengthening). Peak downward movement coincided with cessation of aortic flow and diastolic lengthening with flow through the atrioventricular valves. Long-axis function in the fetus is feasible if M-mode angle correction is used. There was no clinically significant difference between tricuspid and mitral valve excursions. The pattern of atrioventricular valve movement is coordinate and similar to that of adults. Long-axis function offers a new avenue to study systolic and diastolic function in the fetus.


Nuclear pores in rat hypothalamic arcuate neurons: sex differences and changes during the oestrous cycle.
The numerical density of nuclear pores was assessed on freeze-fracture replicas of hypothalamic arcuate neurons from adult male and female rats. In females the nuclear pore density fluctuated during the oestrous cycle and was higher in oestrus than in pro-oestrus, metoestrus and dioestrus. Nuclear pore density in males and in metoestrus and dioestrus females was similar. The nuclear pore density in male rats was significantly less than that in oestrus and pro-oestrus females. The variation of the number of pores per unit nuclear volume and the total number of pores per nucleus was similar to the variation of the numerical density of nuclear pores. These results provide morphological evidence of cyclic changes in neuronal nucleocytoplasmic traffic during the ovarian cycle.


Effect of intrafraction prostate motion on proton pencil beam scanning delivery: a quantitative assessment.
To assess the dosimetric impact caused by the interplay between intrafraction prostate motion and the intermittent delivery of proton pencil beam scanning (PBS). A cohort of 10 prostate patients was treated with PBS using a bilateral single-field uniform dose (SFUD) modality. Bilateral intensity-modulated proton therapy (IMPT) plans were generated for comparison. Because beam-on time in PBS was intermittent, the actual beam-on time was determined from treatment logs. Prostate motion was generalized according to real-time Calypso tracking data from our previously reported prospective photon trial. We investigated potential dose deviations by considering the interplay effect resulting from the worst-case scenario motion and the PBS delivery sequence. For both bilateral-field SFUD and IMPT plans, clinical target volume (CTV) D99% coverage was degraded <2% owing to prostate intrafraction motion when averaged over the course of treatment, but was >10% for the worst fraction. The standard deviation of CTV D99% distribution was approximately 1.2%. The CTV coverage of individual fields in SFUD plans degraded as time elapsed after the initial alignment, owing to prostate drift. Intensity-modulated proton therapy and SFUD demonstrated comparable results when bilateral opposed fields were used. Single-field SFUD plans that were repainted twice, which could reduce half of the treatment time, resulted in similar CTV coverage as bilateral-field plans. Intrafraction prostate motion affects the actual delivered dose to CTV; however, when averaged over the course of treatment, CTV D99% coverage degraded only approximately 2% even for the worst-case scenario. The IMPT plan results are comparable to those of the SFUD plan, and similar coverage can be achieved if treated by SFUD 1 lateral field per day when rescanning the field twice to shorten the treatment time and mitigate intrafraction motion.


Effect of voluntary dehydration on thermoregulatory responses to heat in men and women.
The effects of dehydration prior to heat exposure on sweating and body temperature were tested in 8 men and 8 women, dehydration being 1.3 and 1.0% of body weight, respectively. The subjects were exposed to 40 degrees C for 60 min. Compared with controls (C), in the dehydrated men (D) there was a longer delay in the onset of sweating (C, 7.8, D, 11.6 min, p less than 0.05), a lower total sweat loss (C, 153, D, 127 g X m-2 X h-1, p less than 0.001), and a greater increase in Tre (C, 0.31, D, 0.43 degree C, p less than 0.002). In women, dehydration did not influence the control time course of sweating significantly, nor were these significant body temperature increases during heat exposure. Delay in the onset of sweating in women (C, 18.1, D, 18.7 min) was generally longer than in men (C, 7.8, D, 11.6 min), [F(1,14) = 7.41, p less than 0.05]. A significant correlation was found between the inertia time of sweating and delta Tre in both control and dehydration conditions in the men (r = 0.81, p less than 0.01). The rectal temperature increases in men were also related to the inertia time of electrical skin resistance (r = 0.83, p less than 0.01). It is concluded that dehydration affects sweating and body temperature in men more severely than in women.


An engineered ACE2 decoy receptor can be administered by inhalation and potently targets the BA.1 and BA.2 omicron variants of SARS-CoV-2.
Monoclonal antibodies targeting the SARS-CoV-2 spike (S) glycoprotein neutralize infection and are efficacious for the treatment of mild-to-moderate COVID-19. However, SARS-CoV-2 variants have emerged that partially or fully escape monoclonal antibodies in clinical use. Notably, the BA.2 sublineage of B.1.1.529/omicron escapes nearly all monoclonal antibodies currently authorized for therapeutic treatment of COVID-19. Decoy receptors, which are based on soluble forms of the host entry receptor ACE2, are an alternative strategy that broadly bind and block S from SARS-CoV-2 variants and related betacoronaviruses. The high-affinity and catalytically active decoy sACE2 2 .v2.4-IgG1 was previously shown to be effective in vivo against SARS-CoV-2 variants when administered intravenously. Here, the inhalation of sACE2 2 .v2.4-IgG1 is found to increase survival and ameliorate lung injury in K18-hACE2 transgenic mice inoculated with a lethal dose of the virulent P.1/gamma virus. Loss of catalytic activity reduced the decoy's therapeutic efficacy supporting dual mechanisms of action: direct blocking of viral S and turnover of ACE2 substrates associated with lung injury and inflammation. Binding of sACE2 2 .v2.4-IgG1 remained tight to S of BA.1 omicron, despite BA.1 omicron having extensive mutations, and binding exceeded that of four monoclonal antibodies approved for clinical use. BA.1 pseudovirus and authentic virus were neutralized at picomolar concentrations. Finally, tight binding was maintained against S from the BA.2 omicron sublineage, which differs from S of BA.1 by 26 mutations. Overall, the therapeutic potential of sACE2 2 .v2.4-IgG1 is further confirmed by inhalation route and broad neutralization potency persists against increasingly divergent SARS-CoV-2 variants.


Physiological mechanism of increase in deoxy-hemoglobin concentration during neuronal activation in patients with cerebral ischemia: a simulation study with the balloon model.
Patients with cerebral ischemia or brain tumor have been reported to exhibit an increase of deoxygenated hemoglobin (deoxy-Hb) together with an increase of oxygenated hemoglobin (oxy-Hb). However, the physiological mechanisms underlying this hemodynamic response pattern are unclear. In this study, we performed a simulation using the balloon model (Buxton et al., Magn Reson Med 39:855-864, 1998). We hypothesized that the oxygen extraction rate during the rest period (E 0) in the patients is larger than in normal subjects, because the cerebral blood flow and the speed at which the blood passes through the brain tissues are lower in the patients. The simulation result showed an increase of deoxy-Hb as well as oxy-Hb, especially when E 0 is extremely high. Thus, the results of our simulation suggest that the increase of deoxy-Hb during activation in patients with ischemia or brain tumor is caused by an increased oxygen extraction rate at rest, compared with that of healthy adults.


Creation of brachial artery-basilic vein fistula. A supplement to the cavopulmonary shunt.
Since 1984 five patients who had previously had cavopulmonary shunts for cyanotic congenital heart disease have returned to the Birmingham Childrens' Hospital with increasing breathlessness and cyanosis. Two had had a classic Glenn (unidirectional) cavopulmonary shunt and three, a bidirectional shunt; all shunts had been performed 5 to 18 years earlier. Each patient was reinvestigated and underwent creation of a brachial artery-basilic vein fistula. This procedure is known to have good patency rates when used in other situations (for example, for hemodialysis), but it has not been previously described for use in this context. Of the five patients, four had a considerable and sustained symptomatic improvement on follow-up of 12 to 49 months (mean 31 months), but the condition of one continued to deteriorate; that child has subsequently undergone a Fontan procedure, also with no improvement. Only one patient complained of coldness of the arm, and there were no other complications. We believe a brachial artery-basilic vein fistula has considerable advantages in terms of ease of operation and postoperative complications when compared with an axillary arteriovenous fistula, used as a supplement to an inadequate cavopulmonary shunt.


Use of a combination of an anti-inflammatory steroid, an antibacterial agent and an antifungal in dermatological practice.
Forty patients with skin diseases of various origins were treated with a combination of three creams, the respective bases of which were clobetasone butyrate, sodium fusidate and ketoconazole. Satisfactory results were obtained in 97.5% of cases, with remission of symptoms and good healing of lesions. Tolerance was excellent in 97.5% of cases, with only one patient complaining of burning sensations after the applications.


Divergences in the response to ultraviolet radiation between polar and non-polar ciliated protozoa: UV radiation effects in Euplotes.
Ultraviolet (UV) radiation has detrimental effects on marine ecosystems, in particular in the polar regions where stratospheric ozone reduction causes higher levels of solar radiation. We analyzed two polar species of Euplotes, Euplotes focardii and Euplotes nobilii, for the sensitivity to UV radiation in comparison with two akin species from mid-latitude and tropical waters. Results showed that they face UV radiation much more efficiently than the non-polar species by adopting alternative strategies that most likely reflect different times of colonization of the polar waters. While E. focardii, which is endemic to the Antarctic, survives for longer exposed to UV radiation, E. nobilii, which inhabits both the Antarctic and Arctic, recovers faster from UV-induced damage.