Prevalence of AAV2.5 neutralizing antibodies in synovial fluid and serum of patients with osteoarthritis.
Osteoarthritis (OA) is a leading cause of disability with no cure and only supportive therapy. Adeno-associated virus (AAV) serotype 2.5 is being used in a Phase I clinical trial to deliver the interleukin-1 receptor antagonist into knee joints with OA. Neutralizing antibodies (Nab) directed against AAV2.5, if present, could inhibit gene transfer. Here, we report the prevalence of AAV2.5 Nab in the sera and synovial fluids of patients with OA. Nab titers were measured by their ability to inhibit in vitro transduction by AAV2.5 encoding GFP. Of 44 synovial fluids from patients with mid-stage and advanced OA, 43% had undetectable Nab; 25% had low titers (<1:100), 16% had medium titers (1:100-1:1000) and 16% had high titers (>1:1000) of Nab. Titers of AAV2.5 Nabs correlated with those of AAV2, but not with those of AAV5. Serum titers of AAV2.5 Nab correlated positively with titers in synovial fluid, and were never less than the matched synovial fluid titers. These findings suggest that high titers of Nab against AAV2.5 are uncommon in the synovial fluids of patients with OA, and individuals with high synovial fluid Nab titers can be identified by measuring titers in the serum.


Effects of antiglucocorticoid RU 486 on development of obesity in obese fa/fa Zucker rats.
The effects of RU 486 (mitepristone), an antagonist of type II glucocorticoid receptors (GR), on the development of obesity in young 5-wk-old obese fa/fa rats has been investigated. After 15 days of treatment, body composition of obese RU 486-treated rats was similar to that of lean-vehicle rats. Analysis of body composition changes showed that RU 486 effectively reversed the obesity. It stopped fat deposition in obese rats but increased protein deposition to the level of lean-vehicle rats. RU 486 prevented the development of hyperphagia and reduced gross energetic efficiency in the obese rats but had little effect on lean rats. Brown adipose tissue mitochondrial GDP binding was increased in obese rats but was reduced in lean rats by RU 486 treatment. RU 486 also reduced the elevated activity of hippocampal glycerophosphate dehydrogenase, a glucocorticoid-responsive enzyme, of obese rats to the level of lean rats. The evidence suggests that abnormal activity of glucocorticoid GR receptors or abnormal cellular responsiveness to corticosterone receptor complexes may be important in the development of obesity in the fa/fa rat.


Bee venom improves diabetic wound healing by protecting functional macrophages from apoptosis and enhancing Nrf2, Ang-1 and Tie-2 signaling.
Impaired wound healing is a serious complication of diabetes that negatively affects the patient's socioeconomic life. Multiple mechanisms contribute to impaired diabetic wound healing including deficient recruitment of wound macrophages/neutrophils and impaired neovascularization. Bee venom (BV) has been used as an anti-inflammatory agent for the treatment of several diseases. Nevertheless, the impacts of BV on the diabetic wound healing have been poorly studied. In the present study, we investigated the molecular mechanisms underlying BV treatment on diabetic wound healing in a type I diabetic mouse model. Three experimental groups were used: group 1, non-diabetic control mice; group 2, vehicle-diabetic mice; and group 3, BV-treated diabetic mice. We found that the diabetic mice exhibited impaired wound closure characterized by a significant decrease in collagen and β-defensin-2 (BD-2) expression compared to control non-diabetic mice. The impairment of diabetic wound healing is attributed to increased ROS levels and abolished antioxidant enzymes activity in the wounded tissues. Additionally, wounded tissue in diabetic mice revealed aberrantly decreased levels of Ang-1 and Nrf2 (the agonist ligands of Tie-2) followed by a marked reduction in the phosphorylation of Tie2 and downstream signaling eNOS, AKT and ERK. Impaired diabetic wound healing was also characterized by a significant reduction in activities of total antioxidant enzymes followed by a marked reduction in the levels of CCL2, CCL3 and CXCL2; which led to impaired recruitment and functions of wound macrophages/neutrophils; and significant reduction in the expression of CD31, a marker for neovascularization and angiogenesis of the injured tissue. Interestingly, BV treatment significantly enhanced wound closure in diabetic mice by increasing collagen and BD-2 expression and restoring the levels of Ang-1 and Nrf2 and hence enhancing the Tie-2 downstream signaling. Most importantly, treatment of diabetic mice with BV significantly restored the activities of wounded tissue antioxidant enzymes and the levels of chemokines, and subsequently rescued wound macrophages from mitochondrial membrane potential-induced apoptosis. Our findings reveal the immune-enhancing effects of BV for improving healing process of diabetic wounds and provide the first insight concerning the underlying molecular mechanisms.


Actin Cytoskeleton Role in the Maintenance of Neuronal Morphology and Long-Term Memory.
Evidence indicates that long-term memory formation creates long-lasting changes in neuronal morphology within a specific neuronal network that forms the memory trace. Dendritic spines, which include most of the excitatory synapses in excitatory neurons, are formed or eliminated by learning. These changes may be long-lasting and correlate with memory strength. Moreover, learning-induced changes in the morphology of existing spines can also contribute to the formation of the neuronal network that underlies memory. Altering spines morphology after memory consolidation can erase memory. These observations strongly suggest that learning-induced spines modifications can constitute the changes in synaptic connectivity within the neuronal network that form memory and that stabilization of this network maintains long-term memory. The formation and elimination of spines and other finer morphological changes in spines are mediated by the actin cytoskeleton. The actin cytoskeleton forms networks within the spine that support its structure. Therefore, it is believed that the actin cytoskeleton mediates spine morphogenesis induced by learning. Any long-lasting changes in the spine morphology induced by learning require the preservation of the spine actin cytoskeleton network to support and stabilize the spine new structure. However, the actin cytoskeleton is highly dynamic, and the turnover of actin and its regulatory proteins that determine and support the actin cytoskeleton network structure is relatively fast. Molecular models, suggested here, describe ways to overcome the dynamic nature of the actin cytoskeleton and the fast protein turnover and to support an enduring actin cytoskeleton network within the spines, spines stability and long-term memory. These models are based on long-lasting changes in actin regulatory proteins concentrations within the spine or the formation of a long-lasting scaffold and the ability for its recurring rebuilding within the spine. The persistence of the actin cytoskeleton network within the spine is suggested to support long-lasting spine structure and the maintenance of long-term memory.


Gene conversions and their relation to homologous chromosome pairing.
Gene conversion is the non-reciprocal transfer of DNA sequences from one gene to a related gene elsewhere in the genome. Molecular evidence for its occurrence in higher eukaryotes was first described by our laboratory in 1980 in the two linked human foetal gamma-globin genes. Over a kilobase of DNA was converted in this initial example. Other investigators have since described more examples of gene conversion including some in which the sequence that was transferred is much shorter. We have now accumulated evidence for a series of such small gene conversions in the human foetal globin gene pair. The number of small gene conversions that we have been able to detect leads us to suggest that gene conversions are the consequence of a general mechanism whereby DNA strand invasions enable chromosomes to find their homologues during meiosis.


[Progenitors of transitory spleen colonies in mouse embryonal liver].
The transitory nature of about half of spleen colonies macroscopically detectable in the spleen 7 days after injection of embryonal liver hemopoietic cells was demonstrated by localization of the colonies on the spleen surface and by the study of the content of polypotential and unipotential precursors in individual 7- and 11-day colonies produced in the spleen of irradiated mice by the cells of early (12-13-day) and late (17-18-day) embryonal liver.


SPM-1-producing Pseudomonas aeruginosa ST277 clone recovered from microbiota of migratory birds.
The production of São Paulo metallo-β-lactamase (SPM-1) is the most common carbapenem resistance mechanism detected among multidrug-resistant Pseudomonas aeruginosa clinical isolates in Brazil. Dissemination of SPM-1-producing P. aeruginosa has been restricted to the nosocomial settings, with sporadic reports of environmental isolates due to contamination by hospital sewage. Herein, we described the detection and molecular characterization of SPM-1-producing P. aeruginosa recovered from the microbiota of migratory birds in Brazil. Three hundred gram-negative bacilli were recovered from cloacal and choanal swabs of Dendrocygna viduata during a surveillance study for detection of carbapenem-resistant isolates. All isolates were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. MICs were determined by agar dilution, except for polymyxin B. Antibiotic resistance genes were detected by polymerase chain reaction (PCR) followed by DNA sequencing. Transcriptional levels of oprD and efflux system encoding genes were also carried out by quantitative real-time PCR. Nine imipenem-resistant P. aeruginosa isolates were recovered with 7 of them carrying blaSPM-1. Additional resistance genes (rmtD-1, blaOXA-56,aacA4, and aac(6')-Ib-cr) were also detected in all 9 isolates. The SPM-1-producing isolates showed high MICs for all β-lactams, fluoroquinolones, and aminoglycosides, being susceptible only to polymyxin B. Interestingly, all isolates showed the same PFGE pattern and belonged to ST277. Overexpression of MexXY-OprM and MexAB-OprM was observed in those isolates that did not harbor blaSPM-1. Our results suggest that migratory birds might have played a role in the dissemination of SPM-1-producing P. aeruginosa within the Brazilian territory.


Two unusual hepatitis C virus subtypes, 2j and 2q, in Spain: Identification by nested-PCR and sequencing of a NS5B region.
Many studies have reported the use of the NS5B gene to subtype hepatitis C virus (HCV). Other HCV genes, such as HCV-5' UTR, Core (C) and E1, have also been used. In some studies, NS5B have been used together with 5'-UTR or C genes to improve genotyping results obtained using commercial procedures. Only two studies in Spain have compared molecular techniques versus commercial procedures regarding the efficacy of HCV subtyping. The aim of this study was to determine whether nested PCR and sequencing of a NS5B region was more reliable than commercial procedures to subtype HCV. We analyzed the results of HCV genotyping in [726] serum specimens collected from 2001 to 2013. From 2001 to 2011, we used PCR and INNO-LiPA hybridization or its new version Versant HCV Genotype 2.0 assay (471 samples). From 2012 to 2013, we used nested PCR and sequencing of a NS5B region (255 cases). This method used two pairs of primers to amplify the RNA of the sample converted to DNA by retrotranscription. The amplification product of 270 base pairs was further sequenced. To identify the subtype, the sequences obtained were compared to those in the international database: http://hcv.lanl.gov./content/sequence/, HCV/ToolsOutline.html and Geno2pheno[hcv] http://hcv.bioinf.mpi-inf.mpg.de/index.php. Nested PCR of a NS5B region and sequencing identified all but one subtype (0.4%, 1/255), differentiated all 1a subtypes from 1b subtypes, and characterized all HCV 2-4 subtypes. This approach also distinguished two subtypes, 2j and 2q, that had rarely been detected previously in Spain. However, commercial procedures failed to subtype 12.7% (60/471) of samples and to genotype 0.6% of specimens (3/471). Nested PCR and sequencing of a NS5B region improved the subtyping of HCV in comparison with classical procedures and identified two rare subtypes in Spain: 2j and 2q. However, full length genome sequencing is recommended to confirm HCV 2j and 2q subtypes.


Identification of Neuronal Pentraxins as Synaptic Binding Partners of C1q and the Involvement of NP1 in Synaptic Pruning in Adult Mice.
Elements of the immune system particularly that of innate immunity, play important roles beyond their traditional tasks in host defense, including manifold roles in the nervous system. Complement-mediated synaptic pruning is essential in the developing and healthy functioning brain and becomes aberrant in neurodegenerative disorders. C1q, component of the classical complement pathway, plays a central role in tagging synapses for elimination; however, the underlying molecular mechanisms and interaction partners are mostly unknown. Neuronal pentraxins (NPs) are involved in synapse formation and plasticity, moreover, NP1 contributes to cell death and neurodegeneration under adverse conditions. Here, we investigated the potential interaction between C1q and NPs, and its role in microglial phagocytosis of synapses in adult mice. We verified in vitro that NPs interact with C1q, as well as activate the complement system. Flow cytometry, immunostaining and co-immunoprecipitation showed that synapse-bound C1q colocalizes and interacts with NPs. High-resolution confocal microscopy revealed that microglia-surrounded C1q-tagged synapses are NP1 positive. We have also observed the synaptic occurrence of C4 suggesting that activation of the classical pathway cannot be ruled out in synaptic plasticity in healthy adult animals. In summary, our results indicate that NPs play a regulatory role in the synaptic function of C1q. Whether this role can be intensified upon pathological conditions, such as in Alzheimer's disease, is to be disclosed.


A controlled study of depression among attendees of an oncology clinic in West Africa.
Depression is commonly encountered among cancer patients and has been linked with grave consequences if not promptly treated. This study was set to determine the prevalence of depression in cancer patients. The samples were recruited using systematic random sampling and made up of 200 cancer patients and 200 age and sex matched controls that consented and met the inclusion criteria. The instruments used included: Socio-demographic and clinical profile questionnaire, Centre for Epidemiological Studies Depression Scale Revised (CES-DR), and the Schedule for Clinical Assessment in Neuropsychiatry (SCAN). The instruments were administered by the researchers on the subjects and controls, and data obtained were analyzed using SPSS-15. A larger proportion of cancer patients compared to controls had depressive symptomatology (CES-DR score of 16 and above) made up of 98 (49.0%) cancer patients compared to 40 (20.0%) controls, and the difference was statistically significant, p = < 0.001. Seventy-four (18.5%) subjects from the two groups had depressive disorder using SCAN. The prevalence of depression in cancer subjects was 27.5% with 9.5% in controls and the difference was statistically significant with p = < 0.001. A substantial proportion of cancer patients with depression, 36 (65.5%, n = 55), had moderate depression, 15 (27.3%, n = 55) were mildly depressed, and 7.2% (n = 55) had severe depression. A significant proportion of cancer patients had depression compared with the controls with acute medical conditions. Prompt identification and treatment of cancer, mental disorders like depression should be integrated into cancer care in this part of the world.


Interactions of individual perceived barriers and neighbourhood destinations with obesity-related behaviours in Europe.
Perceived barriers towards physical activity and healthy eating as well as local availability of opportunities (destinations in the neighbourhood) are important determinants of obesity-related behaviours in adults. Little is known, however, about how these factors interact with the behaviours. Data were analysed from 5,205 participants of the SPOTLIGHT survey, conducted in 60 neighbourhoods in urban regions of five different countries across Europe. A virtual audit was conducted to collect data on the presence of destinations in each neighbourhood. Direct associations of, and interactions between, the number of individual perceived barriers and presence of destinations with obesity-related behaviours (physical activity and dietary behaviours) were analysed using multilevel regression analyses, adjusted for key covariates. Perceiving more individual barriers towards physical activity and healthy eating was associated with lower odds of physical activity and healthy eating. The presence of destinations such as bicycle lanes, parks and supermarkets was associated with higher levels of physical activity and healthier dietary behaviours. Analyses of additive interaction terms suggested that the interaction of destinations and barriers was competitive, such that the presence of destinations influenced obesity-related behaviours most among those perceiving more barriers. These explorative findings emphasize the interest and importance of combining objective (e.g. virtual neighbourhood audit) methods and subjective (e.g. individual perceived barriers collected in a survey) to better understand how the characteristics of the residential built environment can shape obesity-related behaviours depending on individual characteristics.


Budesonide/formoterol combination therapy as both maintenance and reliever medication in asthma.
Asthma control is improved by combining inhaled corticosteroids with long-acting beta2-agonists. However, fluctuating asthma control still occurs. We hypothesized that in patients receiving low maintenance dose budesonide/formoterol (bud/form), replacing short-acting beta2-agonist (SABA) reliever with as-needed bud/form would provide rapid symptom relief and simultaneous adjustment in antiinflammatory therapy, thereby reducing exacerbations. In this double-blind, randomized, parallel-group study, 2,760 patients with asthma aged 4-80 years (FEV1 60-100% predicted) received either terbutaline 0.4 mg as SABA with bud/form 80/4.5 microg twice a day (bud/form + SABA) or bud 320 microg twice a day (bud + SABA) or bud/form 80/4.5 microg twice a day with 80/4.5 microg as-needed (bud/form maintenance + relief). Children used a once-nocte maintenance dose. Bud/form maintenance + relief prolonged time to first severe exacerbation (p < 0.001; primary endpoint), resulting in a 45-47% lower exacerbation risk versus bud/form + SABA (hazard ratio, 0.55; 95% confidence interval, 0.44, 0.67) or bud + SABA (hazard ratio, 0.53; 95% confidence interval 0.43, 0.65). Bud/form maintenance + relief also prolonged the time to the first, second, and third exacerbation requiring medical intervention (p < 0.001), reduced severe exacerbation rate, and improved symptoms, awakenings, and lung function compared with both fixed dosing regimens.


[Resistance of small strongyles in an equine stud in South Africa to the benzimidazole anthelmintics].
This paper is apparently the first report of resistance of helminths of equids to anthelmintics in South Africa. While a strain of Cyathostominae from an Arab horse stud near Pretoria showed greatly reduced susceptibility to benzimidazoles in faecal egg reduction tests, ivermectin was apparently still unaffected. It is suggested that integrated methods of control such as alternation of horses and ruminants on pasture and mechanical removal of faeces should be used in addition to anthelmintics.


CpG oligodeoxynucleotides stimulate IFN-gamma-inducible protein-10 production in human B cells.
Several classes of CpG oligodeoxynucleotides (ODNs) with different immune stimulatory profiles were recently identified: the A-, B- and C-classes. In this study, we investigated the CpG-dependent stimulation of IFN-gamma-inducible protein 10 (IP-10 or CXCL10) in different human immune cell types. CpG ODNs induced IP-10 in monocytes, pDCs and in B cells. Purified B cells as well as RPMI 8226 cells responded to CpG stimulation by IP-10 production. Treatment with exogenous IFN-alpha2b sensitized PBMCs, purified B cells as well as RPMI 8226 cells to respond more efficiently to stimulation with CpG ODNs by IP-10 production. IP-10 signaling could be directly stimulated via TLR9 in CpG-unresponsive HEK293 cells transfected with human TLR9 and an IP-10 reporter construct. Therefore, CpG-mediated IP-10 production is stimulated through IFN-alpha in cells that express the IFN-alpha receptor, a second pathway for IP-10 induction exists in TLR9-expressing B cells and pDCs where IP-10 is stimulated directly upon CpG-mediated TLR9 signaling. Our data provide a better understanding of the mechanisms through which CpG ODNs induce efficient Th1 responses.


Leukocyte telomere length pattern in a Chuvash population that experienced mass famine in 1922-1923: a retrospective cohort study.
To our knowledge, there are no experimental studies that have addressed the effects of starvation on the maintenance of telomere length. Two epidemiologic studies that have addressed this topic gave controversial results. We characterized leukocyte telomere length (LTL) in a Chuvash population that was comprised of survivors of the mass famine of 1922-1923 and in these survivors' descendants. The tested cohort consisted of native Chuvash men (n = 687) and women (n = 647) who were born between 1909 and 1980 and who resided in small villages in the Chuvash Republic of the Russian Federation. Data were gathered during 3 expeditions undertaken in 1994, 1999, and 2002. With the use of this method of gathering the study cohort, we were able to treat age and birth year as independent variables (i.e., after adjustment for age, we were able to analyze how LTL correlates with a birth year in the interval between 1909 and 1980). The DNA of peripheral blood leukocytes was used to measure the telomere length with a quantitative polymerase chain reaction technique. The main observations were as follows: 1) there were shorter leukocyte telomeres in men born after 1923 (i.e., after the mass famine) than in men born before 1922 (i.e., before the mass famine); 2) there was a stable inheritance of shorter telomeres by men of ensuing generations; and 3) there was an absence of a correlation between LTL and birth year in women. Our study does not provide direct evidence for leukocyte telomere shortening in famine survivors. However, the comparative analysis of LTL in the survivors and their descendants suggests that such an effect did take place. The study also implies that mass famine may be associated with telomere shortening in male descendants of famine survivors. This observation is in agreement with the "thrifty telomere hypothesis" predicting that longer telomeres are disadvantageous in nutritionally marginal environments.


Low back pain, smoking and employment during pregnancy and after delivery - a 3-month follow-up study.
Low back pain (LBP.), smoking and employment was studied among 111 consecutive women admitted to a maternity ward over a 6-week period, 40 were primiparas and 71 multiparas. LBP was defined as any pain in the low back, irrespective of the specific cause of the pain. Two specially constructed questionnaires were utilised. The first was a, 14-item questionnaire which all participants answered before leaving the maternity ward. It included questions on employment and smoking and self-rating Visual Analogue Scales used for rating LBP. LBP was rated during the pregnancy and 3 days after delivery. The second questionnaire was used in a 90-day follow-up interview. The mean age of participants was 28 years. The prevalence of LBP during pregnancy was 58.5% among the 111 participants. Of the 111, 75% continued to have LBP postpartum and at the 90-day post-delivery follow-up, 54% of those with LBP during pregnancy were still experiencing LBP. Previous births and birth weight were not found to correlate positively with LBP. LBP during pregnancy did not affect the length of employment during pregnancy. Smokers had LBP more frequently during pregnancy and also after (P <0.002). It is concluded that smoking does seem to contribute to LBP during and after pregnancy. Birth weight does not affect LBP and LBP does not affect the length of employment during pregnancy.


Social complexity in chinese coastal neolithic sites.
Neolithic cemeteries in Zhejiang, Jiangsu, and Shandong provinces from about 5000 to 2000 B.C. show increases in the mean numbers of burial goods, goods found with males, and tools used as grave goods; separation of burial localities; differentiation of sets of tools found with males and females; a decrease in goods found with children; and common ornaments found with males and females. The distribution of tools seems to reflect a sexual division, with an increase in the prominence of males which may be associated with intensification of cultivation. Emergence of ranking is suggested by very rich graves and ornaments shared by males and females. Chinese theories of cultural evolution are introduced.


Fenestrated endovascular aortic aneurysm repair using physician-modified endovascular grafts versus company-manufactured devices.
Fenestrated endografts are customized, patient-specific endovascular devices with potential to reduce morbidity and mortality of complex aortic aneurysm repair. With approval from the U.S. Food and Drug Administration, our center began performing fenestrated endovascular aneurysm repair through a physician-sponsored investigational device exemption (IDE #G130210), using both physician-modified endografts (PMEGs) and company-manufactured devices (CMDs). Because these techniques are associated with specific advantages and disadvantages, we sought to investigate differences in outcomes between PMEG and CMD cases. A single-institution retrospective review of all fenestrated endovascular aneurysm repairs was performed. The cohort was analyzed by device type (PMEG or CMD) after matching of cases on the basis of (1) number of target vessels intended for treatment, (2) extent of aneurysm, (3) aneurysm diameter, (4) device configuration, and (5) date of operation. Outcomes of ruptures, common iliac artery aneurysms, and aortic arch aneurysms were excluded. Demographics, operative details, perioperative complications, length of stay, and reinterventions were compared. For patients with >1 year of follow-up time, survival, type I or type III endoleak rate, target artery patency, and reintervention rate were estimated using the Kaplan-Meier method. Between November 30, 2010, and July 30, 2016, 82 patients were identified and matched. The cohort included 41 PMEG and 41 CMD patients who underwent repair of 38 juxtarenal (PMEG, 17; CMD, 21; P = .38), 14 pararenal (PMEG, 6; CMD, 8; P = .56), and 30 thoracoabdominal type I to type IV (PMEG, 18; CMD, 12; P = .17) aneurysms. There were significant differences in presentation requiring urgent aneurysm repair (PMEG, 9; CMD, 0; P = .002), total fluoroscopy time (PMEG, 76 minutes; CMD, 61 minutes; P = .02), volume of contrast material used (PMEG, 88 mL; CMD, 70 mL; P = .02), in-operating room to out-of-operating room time (PMEG, 391 minutes; CMD, 319 minutes; P = .001), incision to surgery end time (PMEG, 276 minutes; CMD, 224 minutes; P = .002), and 1-year reintervention rate (PMEG, 37%; CMD, 13%; log-rank P = .04). No differences in perioperative complications, overall length of stay, type I or type III endoleak, or survival were observed between PMEG and CMD. For the entire cohort including both PMEG and CMD, the overall rate of any 30-day postoperative complication was 39%, and the Kaplan-Meier estimate of survival at 1 year was 86%. In this single-institution experience of fenestrated endovascular aneurysm repair, the primary differences between PMEG and CMD related only to operative metrics and the need for postoperative reinterventions. No statistically significant advantage was found for one approach over the other; we therefore cannot conclude that one approach is better than the other. Both remain viable options that may compare favorably with open repair of complex aortic aneurysms. Further studies are necessary to determine whether this relative equivalence represents a type II error or lack of long-term durability data or whether true equivalence between PMEG and CMD approaches exists.


[The special training month for tutors and residents in family medicine in Valencia: evaluation of 3 cases].
To evaluate the activity in the special training month (STM) for tutors and residents in Family Medicine in Valencia between 1994 and 1996. Descriptive and longitudinal. Thirteen teaching health centres. 48 tutors and 102 residents. Requests from tutors for clinical recycling, research and other activities. Evaluations of hospital staff, tutors and residents. 41.6% of tutors did their STM in three years, 29.2% in 2 years and 29.2% in one. Of the 105 evaluations sent in, 92 (87.6%) were for clinical recycling, 9 (8.6%) research and 4 (3.8%) other activities. The most visited specialities were Dermatology, Cardiology, Radiology and Rheumatology. Staff evaluated as very good the interest shown and punctuality; between good and very good, their knowledge and skills, with commentaries in 36% of evaluations on improvement of coordination between levels. 64.4% of tutors considered the STM very favourably, reflected in nine commentaries that the activity was motivating and in six that it promoted relationships between the Primary and Specialist care levels; and that it had an objective repercussion on teaching through 44 clinical sessions reported during 1996. Improvements were proposed in the organisation of teaching activities. 49% of residents were assigned a set tutor. The STM is valued positively by tutors, residents and staff.


Inhibitory effect of methyl gallate and gallic acid on oral bacteria.
This study examined the ability of methyl gallate (MG) and gallic acid (GA), the main compounds of gallo-tannins in Galla Rhois, to inhibit the proliferation of oral bacterial and the in vitro formation of Streptococcus mutans biofilms. The antimicrobial activities of these compounds were evaluated in vitro using the broth microdilution method and a beaker-wire test. Both MG and GA had inhibitory effects on the growth of cariogenic (MIC<8 mg/ml) and periodontopathic bacteria (MIC=1 mg/ml). Moreover, these compounds significantly inhibited the in vitro formation of S. mutans biofilms (MG, 1 mg/ml; GA, 4 mg/ml; P<0.05). MG was more effective in inhibiting bacterial growth and the formation of S. mutans biofilm than GA. In conclusion, MG and GA can inhibit the growth of oral pathogens and S. mutans biofilm formation, and may be used to prevent the formation of oral biofilms.


Identification of HDAC10, a novel class II human histone deacetylase containing a leucine-rich domain.
Histone acetylation is important for regulating chromatin structure and gene expression. Three classes of mammalian histone deacetylases have been identified. Among class II, there are five known members, namely HDAC4, HDAC5, HDAC6, HDAC7 and HDAC9. Here we describe the identification and characterization of a novel class II member termed HDAC10. It is a 669 residue polypeptide with a bipartite modular structure consisting of an N-terminal Hda1p-related putative deacetylase domain and a C-terminal leucine-rich domain. HDAC10 is widely expressed in adult human tissues and cultured mammalian cells. It is enriched in the cytoplasm and this enrichment is not sensitive to leptomycin B, a specific inhibitor known to block the nuclear export of other class II members. The leucine-rich domain of HDAC10 is responsible for its cytoplasmic enrichment. Recombinant HDAC10 protein possesses histone deacetylase activity, which is sensitive to trichostatin A, a specific inhibitor for known class I and class II histone deacetylases. When tethered to a promoter, HDAC10 is able to repress transcription. Furthermore, HDAC10 interacts with HDAC3 but not with HDAC4 or HDAC6. These results indicate that HDAC10 is a novel class II histone deacetylase possessing a unique leucine-rich domain.


Organizational cooperation in teleradiology.
Teleradiology requires cooperation between participating parties, but it cannot be assumed that such cooperation will work well. A survey was carried out of 12 radiology departments in Norway which used picture archiving and communication systems in connection with teleradiology. Qualitative interviews were carried out with 26 out of 29 resource persons (response rate 90%). The respondents identified 17 issues of importance for cooperation in teleradiology. None of the problems with cooperation seem large enough to prevent effective teleradiology cooperation in future. For organizations planning teleradiology of a larger volume, the cooperation issue is important. It is recommended that managers lead change processes in their organizations where the different issues of importance to cooperation are treated and the right measures are taken to realize the full potential of teleradiology. For telemedicine, it is important that future research includes investigations on cooperation for the different applications.


Clock gene expression in the human pituitary gland.
Pituitary function relies on strictly timed, yet plastic mechanisms, particularly with respect to the daytime-dependent coordination of hormone synthesis and release. In other systems, clock genes and their protein products are well-described candidates to anticipate the daily demands in neuroendocrine coupling and to manage cellular adaptation on changing internal or external circumstances. To elucidate possible mechanisms of time management, a total of 52 human autoptic pituitary glands were allocated to the 4 time-of-day groups, night, dawn, day, and dusk, according to reported time of death. The observed daytime-dependent dynamics in ACTH content supports a postmortem conservation of the premortem condition, and thus, principally validates the investigation of autoptic pituitary glands. Pituitary extracts were investigated for expression of clock genes Per1, Cry1, Clock, and Bmal1 and corresponding protein products. Only the clock gene Per1 showed daytime-dependent differences in quantitative real-time PCR analyses, with decreased levels observed during dusk. Although the overall amount in clock gene protein products PER1, CRY1, and CLOCK did not fluctuate with time of day in human pituitary, an indication for a temporally parallel intracellular translocation of PER1 and CRY1 was detected by immunofluorescence. Presented data suggest that the observed clock gene expression in human pituitary cells does not provide evidence for a functional intrinsic clockwork. It is suggested that clock genes and their protein products may be directly involved in the daytime-dependent regulation and adaptation of hormone synthesis and release and within homeostatic adaptive plasticity.


Gastric pacing for diabetic gastroparesis--does it work?
The management of diabetic gastroparesis resistant to medical therapy is very difficult Gastric electrical stimulation (GES) is a relatively new therapeutic modality which has shown some promise in international trials. It has seen use in four patients in Ireland. Our aim was to determine if GES improved patients' outcomes in terms of duration and cost of inpatient stay and glycaemic control. We reviewed the patients' case notes and calculated the number of days spent as an inpatient with symptomatic gastroparesis pre and post pacemaker, the total cost of these admissions, and patients' average HbA1c pre and post GES. Mean length of stay in the year pre GES was 81.75 days and 62.25 days in the year post GES (p=0.89). There was also no improvement in glycaemic control following GES. GES has been ineffective in improving length of inpatient stay and glycaemic control in our small patient cohort.


Sorption of organic compounds to fresh and field-aged activated carbons in soils and sediments.
Activated carbon (AC) amendment to polluted sediment or soil is an emerging in situ treatment technique that reduces freely dissolved porewater concentrations and subsequently reduces the ecological and human health risk of hydrophobic organic compounds (HOCs). An important question is the capacity of the amended AC after prolonged exposure in the field. To address this issue, sorption of freshly spiked and native HOCs to AC aged under natural field conditions and fresh AC amendments was compared for one soil and two sediments. After 12-32 months of field aging, all AC amendments demonstrated effectiveness for reducing pore water concentrations of both native (30-95%) and spiked (10-90%) HOCs compared to unamended sediment or soil. Values of K(AC) for field-aged AC were lower than freshly added AC for spiked HOCs up to a factor of 10, while the effect was less for native HOCs. The different behavior in sorbing native HOCs compared to freshly spiked HOCs was attributed to differences in the sorption kinetics and degree of competition for sorption sites between the contaminants and pore-clogging natural organic matter. The implications of these findings are that amended AC can still be effective in sorbing additional HOCs some years following amendment in the field. Thus, a certain level of long-term sustainability of this remediation approach is observed, but conclusions for decade-long periods cannot be drawn solely based on the present study.


The Explanatory Role of Insomnia in the Relationship between Pain Intensity and Posttraumatic Stress Symptom Severity among Trauma-Exposed Latinos in a Federally Qualified Health Center.
Latinos, one of the fastest growing populations in the United States, suffer from high rates of posttraumatic stress symptoms (PTS) and its clinical correlates (e.g., disability). Although research suggests the experience of pain is closely related to PTS among trauma-exposed groups, there has been little exploration of the processes that may link pain intensity to greater PTS among trauma-exposed Latinos. The current study explored insomnia, a common problem associated with both pain intensity and PTS, as a mechanism in the association between pain intensity and PTS among trauma-exposed Latinos (N = 208, Mage = 39.39 years, SD = 11.48) attending a Federally Qualified Health Center. Results indicated that insomnia partially explained the relationship between pain intensity and PTS total score (B = 0.25, 95% CI [0.12, 0.43]), as well as re-experiencing (B = 0.09, 95% CI [0.04, 0.17]), avoidance (B = 0.09, 95% CI [0.04, 0.17]), and arousal symptoms (B = 0.10, 95% CI [0.04, 0.17]). Future work is needed to explore the extent to which insomnia accounts for relations between pain and PTS using longitudinal designs to further clarify theoretical health disparity models involving these comorbid conditions.


Open channel block by Ca2+ underlies the voltage dependence of drosophila TRPL channel.
The light-activated channels of Drosophila photoreceptors transient receptor potential (TRP) and TRP-like (TRPL) show voltage-dependent conductance during illumination. Recent studies implied that mammalian members of the TRP family, which belong to the TRPV and TRPM subfamilies, are intrinsically voltage-gated channels. However, it is unclear whether the Drosophila TRPs, which belong to the TRPC subfamily, share the same voltage-dependent gating mechanism. Exploring the voltage dependence of Drosophila TRPL expressed in S2 cells, we found that the voltage dependence of this channel is not an intrinsic property since it became linear upon removal of divalent cations. We further found that Ca(2+) blocked TRPL in a voltage-dependent manner by an open channel block mechanism, which determines the frequency of channel openings and constitutes the sole parameter that underlies its voltage dependence. Whole cell recordings from a Drosophila mutant expressing only TRPL indicated that Ca(2+) block also accounts for the voltage dependence of the native TRPL channels. The open channel block by Ca(2+) that we characterized is a useful mechanism to improve the signal to noise ratio of the response to intense light when virtually all the large conductance TRPL channels are blocked and only the low conductance TRP channels with lower Ca(2+) affinity are active.


An N-terminal deletion mutant of estrogen receptor exhibits increased synergism with upstream activators and enhanced binding to the estrogen response element.
To study the role of the N-terminal region of the estrogen receptor (ER) in transcription activation and in DNA binding, we constructed a mutant of the Xenopus laevis ER which lacks amino acids 1-159 (XER160/586). In transient transfections, XER160/586 exhibited < 10% of the activity of wild-type XER on a synthetic promoter containing two estrogen response elements (EREs). To examine transcriptional synergism by XER and by XER160/586, we determined the activity of promoters containing EREs and binding sites for either the vitellogenin activator, NF1, or AP1 upstream activator protein. For the three promoters transcription by XER was 2.8-fold greater than expected for additive activities, and transcription by XER160/586 was 6.2-fold greater. These data demonstrate that an upstream activator protein bound near the promoter can partially compensate for the loss of the internal N-terminal (AF1) transactivation domain in XER160/586. Using a promoter interference assay to study the intracellular interaction between ER and the estrogen response element, we found that XER160/586 exhibited a significant increase in affinity for the ERE. Its low basal activity and enhanced affinity for the ERE make XER160/586 an effective dominant negative mutant. When co-expressed with wild-type XER at 1:1 and 5:1 ratios, XER160/586 suppressed the activity of wild-type XER by 57% and > 80%, respectively.


[The possibilities of surgical treatment of hepatocellular cancer in patients with liver cirrhosis].
154 radical liver resections were performed on the reason of the hepatocellular cancer, of them 33% in patients with liver cirrhosis. Liver function was assessed using the Child--Pugh score. Liver cirrhosis and extensive liver resections proved to be the independent complication and lethality risk factors. The extensive liver resections were performed in 70%. The postoperative morbidity rate was 44.8%, the lethality was 5.8%. The concomitant liver cirrhosis reliably worsens postoperative complications (p = 0.001) and lethality (p = 0.0001) rates. The long-term treatment results were analyzed. Thus, liver resection proved to be an appropriate treatment for patients with hepatocellular cancer and liver cirrhosis Child--Pugh stage A, but a thorough patient selection is recommended. Liver resection is contraindicated by liver cirrhosis Child--Pugh stage B. The orthotopic liver transplantation is recommended in such cases, considering that Milan criteria are observed. The rest cases should consider the possibilities of radiofrequency ablation, transarterial chemoembolization, chemotherapy.


Structural analysis of customized 3D printed plate for pelvic bone by comparison with conventional plate based on bending process.
Pelvic bone fracture is highly complex, and its anatomical reduction is difficult. Therefore, patient-specific customized plates have been developed using three-dimensional (3D) printing technology and are being increasingly used. In this study, the reduction status in five representative pelvic fracture models was compared between two groups: the 3D printing plate (3DP) group using a patient-specific 3D printed plate after virtual reduction and the conventional plate (CP) group using a conventional plate by manual bending. The 3DP and CP groups included 10 and 5 cases, respectively. The fractured models were reduced virtually and their non-locking metal plates were customized using 3D printing. The process of contouring the conventional plates to fit the contact surface of the bone with the bending tool was conducted by an experienced pelvic bone trauma surgeon. The reduction and fixation achieved using the two different plate groups was compared, and the significance of differences in the results was analyzed using paired t-tests, after verifying the normality of data distribution. The vertex distances between the surface of the bone and the contact surface of the plate were significantly lower in the 3DP group than in the CP group (0.407 ± 0.342 and 2.195 ± 1.643, respectively, P = 0.008). Length and angular variations, which are measurements of the reduction state, were also lower in the 3DP group than in the CP group (length variation: 3.211 ± 2.497 and 5.493 ± 3.609, respectively, P = 0.051; angular variation: 2.958 ± 1.977 and 4.352 ± 1.947, respectively, P = 0.037). The customized 3D printed plate in the virtual reduction model provided a highly accurate reduction of pelvic bone fractures, suggesting that the customized 3D printed plate may help ensure easy and accurate reduction.


The Effect of an Enriched Sport Program on Children's Executive Functions: The ESA Program.
The effects of physical exercise on executive functions (EFs) are well-documented. EFs are involved in daily activities, and their development determines the quality of people's future life, both in terms of mental health and quality of life. The purpose of the current paper is to evaluate the effects of a physical education program, elaborated within the Enriched Sports Activity Program (ESA Program), an Erasmus + Project, on EFs, namely, visuospatial working memory, inhibitory control, cognitive flexibility, and task switching. Data were collected on November 2017 (t1) and May 2018 (t2). At t1, a sample of 357 children from four European countries (Italy, Germany, Lithuania, and Turkey) performed a cognitive test battery made up of Digit Span Forward/Backward, Stroop Task, and Trail Making Test (TMT), whose order was randomized. From November until May, classrooms from the experimental group followed the ESA Program, while classrooms from the control continued with the ordinary physical education class. At t2, children from both experimental and control groups performed again the cognitive battery. The repeated measures ANOVA showed a significant effect of the ESA Program on the TMT B and on Digit Span Backward, but no significant effects were found on Digit Span Forward and Stroop Task. The introduction of a sport program enriched with cognitive stimuli has beneficial effects for children working memory and cognitive flexibility.


[Na-Li countertransport and the diagnosis of hypertension and combined hypertension and renal artery stenosis].
The rate of Na-Li countertransport was studied in inpatients with essential hypertension (n = 59), chronic diffuse glomerulonephritis (n = 30), chronic pyelonephritis (n = 26), renovascular hypertension (n = 15) and in those with associated renovascular hypertension and essential hypertension (n = 4). Multiple regression analysis has demonstrated that age, body weight and blood plasma lipids do not make any significant contribution to dispersion of the counter transport rate. The mean rate of countertransport in patients with essential hypertension turned out much higher than that in patients with secondary hypertensions. Repeated examinations have shown that in every man, the countertransport rate remains unchanged for 1.5 yr. It is not affected either by hypotensive therapy or surgical treatment. In inpatients with secondary hypertension and low rates of countertransport, high arterial pressure (AP) drops after surgical treatment of the kidneys, renal vessels or adrenals. Surgical treatment of patients with secondary hypertension and high rates of countertransport does not lead to any material decrease of AP. It is assumed that the rate of Na-Li countertransport can be used for diagnosing associated secondary hypertensions and essential hypertension and prediction of AP lowering after surgical treatment.


Chlorinated, brominated and fluorinated organic pollutants in African Penguin eggs: 30 years since the previous assessment.
The African Penguin population has drastically declined over the last 100 years. Changes in food availability due to over-fishing and other oceanographic changes seem to be major causes. However, it has also been 30 years since organic pollutants as a potential factor have been assessed. We analysed penguin eggs collected in 2011 and 2012 from two breeding colonies 640 km apart: Robben Island near Cape Town on the Atlantic Ocean coast, and Bird Island near Port Elizabeth on the Indian Ocean coast of South Africa. We quantified organochlorine pesticides, brominated flame retardants, and perfluorinated compounds (PFCs). Compared to 30 years ago, concentrations of ΣDDT have remained about the same or slightly lower, while ΣPCBs declined almost four-fold. The use of DDT in malaria control is unlikely to have contributed. PFCs were detected in all eggs. Indications (non-significant) of eggshell thinning associated with ΣDDT and ΣPCB was found. It seems therefore that the concentrations of measured organic pollutants the African Penguin eggs are not contributing directly to its current demise, but concerns remain about thinner shells and desiccation. Effects of combinations of compounds and newer compounds cannot be excluded, as well as more subtle effects on reproduction, development, and behaviour.


ADHD with Comorbid Bipolar Disorders: A Systematic Review of Neurobiological, Clinical and Pharmacological Aspects Across the Lifespan.
Attention deficit hyperactivity (ADHD) disorder is a neurodevelopmental disorder characterized by inattention, hyperactivity, disruptive behaviour, and impulsivity. Despite considered typical of children for a long time, the persistence of ADHD symptoms in adulthood gained increasing interest during the last decades. Indeed, its diagnosis, albeit controversial, is rarely carried out even because ADHD is often comorbid with several other psychiatric diosrders, in particular with bipolar disorders (BDs), a condition that complicates the clinical picture, assessment and treatment. The aim of this paper was to systematically review the scientific literature on the neurobiological, clinical features and current pharmacological management of ADHD comorbid with BDs across the entire lifespan, with a major focus on the adulthood. The pharmacology of ADHD-BD in adults is still empirical and influenced by the individual experience of the clinicians. Stimulants are endowed of a prompt efficacy and safety, whilst non-stimulants are useful when a substance abuse history is detected, although they require some weeks in order to be fully effective. In any case, an in-depth diagnostic and clinical evaluation of the single individual is mandatory. The comorbidity of ADHD with BD is still a controversial matter, as it is the notion of adult ADHD as a distinct nosological category. Indeed, some findings highlighted the presence of common neurobiological mechanisms and overlapping clinical features, although disagreement does exist. In any case, while expecting to disentangle this crucial question, a correct management of this comorbidity is essential, which requires the co-administration of mood stabilizers. Further controlled clinical studies in large samples of adult ADHD-BD patients appear extremely urgent in order to better define possible therapeutic guidelines, as well as alternative approaches for this potentially invalidating condition.


Effects of processing on onion skin powder added extrudates.
It is possible to enhance the functional properties of extruded products with the inclusion of fruit and vegetable by-products. Onion skin, a rich source of quercetin and fiber, is considered as waste in the industry and can be used as an alternative ingredient to improve the nutritional value of the extruded products. Three levels (3, 6, and 9%) of onion skin powder (OSP) were added to wheat flour and compared with control (0% OSP). The effect of the extrusion process on accessible quercetin, total phenolic content, and antioxidant activity of the samples were investigated. In addition, carbohydrate digestibility analyses were conducted for the products. Mass spectrometry (LC-MS/MS) results indicated that increasing the OSP level increased the quercetin content. The process caused the release of the entrapped quercetin from OSP, which was revealed by significantly higher quercetin levels for the extrudates. Some of the quercetin was lost during in vitro digestion process. Increasing the OSP level increased antioxidant activity and total phenolic contents of the samples. Total phenolic contents decreased significantly after the processing, yet antioxidant activities were not affected. The extruded products showed high amounts of rapidly available glucose (69.5 g/100 g). The OSP enhancement did not change the carbohydrate digestibility of products. The results indicated that the extrusion process could increase the level of accessible bioactive ingredients, and the level of functional compound addition can be optimized further.


Macronutrients modifed dietary intervention in the management of overweight/obese children and adolescents - Systematic review
The prevalence of obesity in both adults and children is rapidly increasing worldwide. Obesity is one of main causes developing chronic diseases and various problems like economic consequence. It can be affected by genetic, environmental, psychological, and socioeconomic factors. Dietary modification is a well-known important factor in weight control. Precisely, dietary macronutrients composition, food selection, and dietary patterns along with energy restriction can affect weight reduction. The intervention programs related to diet and lifestyle modification for obesity are continuously being conducted in children and adolescents. Several studies demonstrated that dietary macronutrients modification affects body composition and metabolic-related markers. On the other hand, hypocaloric diets regardless of macronutrients composition have been reported to be effective and stable for weight loss in obese children. However, there are few intervention studies for the association between dietary macronutrients composition and obesity in Korean children and adolescents. Therefore, this systematic review was conducted to suggest a basic evidence to identify optimal macronutrients composition that can be applied in obesity management in Korean children and adolescents.


Oral and craniofacial morphology of a patient with Larsen syndrome.
A case is reported of a 15 years old Chinese female with Larsen syndrome exhibiting many characteristic facial and skeletal features plus hitherto unreported orofacial anomalies including delayed dental development; hypodontia affecting all the permanent canines, second and third molars, maxillary lateral incisors and second premolars; morphological anomalies of the maxillary first premolars and molars; a class 3 skeletal pattern and reduced upper facial height. Cephalometric radiographs showed the skull bones to have an appearance similar to that of osteopetrosis.


Analysis of risk factors in diabetics resulted from polycystic ovary syndrome in women by EDA analysis and machine learning techniques.
This study discusses the relationship between Polycystic Ovary Syndrome (PCOS) and diabetes in women, which has become increasingly prevalent due to changing lifestyles and environmental factors. The characteristic that distinguishes women with PCOS is hyperandrogenism which results from abnormal ovarian or adrenal function, which leads to the overproduction of androgens. Excessive androgens in women increase the risk of Type 2 diabetes (T2D) and insulin resistance (IR). Nowadays, diabetes affects people of all ages and is linked to factors such as lifestyle, genetics, stress, and aging. Diabetes, the uncontrolled high blood sugar level can potentially harm kidneys, nerves, eyes, and other organs and there is no cure, making it a concerning disease in developing nations. This research tried to submit the evidence through feature-wise correlation analyses between PCOS and diabetes. Hence, this model utilized the Exploratory Data Analysis (EDA) and the Elbow clustering algorithms for the experimental purpose in which the EDA deeply analyzed the features of PCOS and diabetes and recorded a positive correlation of 95%. The Elbow clustering technique is employed for verifying the correlations identified through EDA. Although limited research exists on this specific disease, this work provides potential evidence for the research community by evaluating the clustering results using Silhouette Score, Calinski-Harabasz Index, and Davies-Bouldin Index.


Overcorrection of lumbar lordosis for adult spinal deformity with sagittal imbalance: comparison of radiographic outcomes between overcorrection and undercorrection.
To determine the correlation of the difference between postoperative lumbar lordosis (LL) and ideal LL with the sagittal vertical axis (SVA) at the final follow-up in patients with adult spinal deformity (ASD). Fifty-one patients with degenerative lumbar kyphosis (DLK) (mean age 66.5 years) who underwent surgical correction with a minimum 2-year follow-up were evaluated. Based on the difference between postoperative LL and ideal LL using the Korean version of Legaye's formula, we divided the 51 patients into two groups: overcorrection (degree of postoperative LL > ideal LL) and undercorrection (degree of postoperative LL < ideal LL). Our clinical series of patients comprised 24 in the overcorrection and 27 in the undercorrection group. No significant differences were found in preoperative pelvic incidence (PI 52.6° vs. 57.3°), sacral slope (SS 23.3° vs. 18.3°), LL (-6.9° vs. -2.3°), thoracic kyphosis (TK 4.7° vs. 4.9°) and SVA (140 vs. 139 mm) except pelvic tilt (PT 29.4° vs. 39.0°), between the two groups. All the patients in the overcorrection group and 16 in the undercorrection group achieved postoperative optimal sagittal balance based on SVA ≤ 50 mm. In addition, significant differences in PT (10.5° vs. 26.7°), SS (42.1° vs. 30.6°), LL (-64.3° vs. -37.1°), TK (22.6° vs. 15.8°), and SVA (-1 vs. 41 mm) between the two groups were observed postoperatively. Furthermore, four patients (16.7 %) in the overcorrection group and eight (50 %) in the undercorrection group had sagittal decompensation at the final follow-up. Our results showed that the difference between postoperative LL and ideal LL had a significant correlation with postoperative and final follow-up SVA in our clinical series. Overcorrection of LL is an effective treatment modality to maintain optimal sagittal alignment in patients with DLK; this suggests that it should be considered in preoperative planning for patients with ASD with sagittal imbalance.


[The role of cyclic nucleotides in reparative bone formation].
Distinct alterations in content of cyclic nucleotides were observed during treatment of 133 patients with fractures of long tubular bones. The following effects were found after initial increase in cAMP content: increase in tissue permeability, aseptic inflammation, inhibition of cell mitosis, acceleration of cell enzymes synthesis, of protein biosynthesis and of the rate of microcirculation in the traumatized region. Elevation of cGMP concentration led to initiation of bone marrow proliferation, to augmented formation of organic matrix and its mineralization. Monitoring of cyclic nucleotides concentration enabled to evaluate the reparative osteogenesis activity.


Acts, omissions, intentions and motives: a philosophical examination of the moral distinction between killing and letting die.
Health care professionals frequently justify moral decisions by appealing to the acts and omissions distinction and the principle of double effect. These principles are often quoted and criticised in the nursing literature, particularly in relation to active and passive euthanasia. Authors are, however, inclined to acknowledge the fact that they are a philosophical minefield, but fall short of providing a substantial critical analysis. Those who uphold the acts and omissions distinction claim that we are morally less responsible for our omissions than for our acts. The principle of double effect suggests that there is a moral distinction between direct and indirect intention and that we are morally less responsible for that which we intend indirectly. This article offers an examination of these often quoted, but sometimes little understood, principles and it is hoped that it will serve as a basis for debate. Since the intention is to challenge assumptions which can lull us into believing that our moral decisions can be justified by appealing to principles such as these, it is necessary to examine them critically and to go beyond a superficial grasp of the implications of such distinctions. There is a tendency to appeal to the acts and omissions doctrine and the principle of double effect in life and death situations such as euthanasia, infanticide and abortion, but are they valuable tools which should be used in moral decision making, or are they so flawed that they can seriously mislead us when it comes to hard cases? The conclusion reached in this paper is that there is no significant moral difference between an act and an omission, particularly within the context of killing and letting die, and that the principle of double effect encourages hypocrisy rather than honesty.


Who are borderline patients?
"Borderline" is a term used with increasing frequency to describe certain difficult patients. It refers to a heterogeneous population of patients who tend to get involved in impaired doctor-patient relationships. Different areas of psychiatry have their own, often conflicting definitions of these disorders. Early trauma, neglect and genetic diatheses to axis I disorders commonly occur in the backgrounds of these patients. Most have a combination of factors. With the multiple different definitions and the potential for misuse of the term, it should be used with caution and with an understanding of what definition is being used. Treatment and prognostic implications need to be individualized.


An Updated Checklist of the Sicilian Native Edible Plants: Preserving the Traditional Ecological Knowledge of Century-Old Agro-Pastoral Landscapes.
The traditional use of native wild food plants (NWFP) may represent a valuable supplementary food source for the present and future generations. In Sicily, the use of wild plants in the human diet dates back to very ancient times and still plays an important role in some rural communities. Moreover, in this regard, the natural and cultural inheritance of this island is wealthy and diversified for several reasons. First, Sicily hosts a rich vascular flora, with 3,000 native and 350 endemic plants. Second, due to its central position in the Mediterranean, the island has acted as a veritable melting pot for the ethnobotanical knowledge of the rural communities of the entire basin. We reviewed all the available literature and, starting from such omnicomprehensive checklist, partially improved thanks to the data issuing from recent field investigations, we critically revised the whole species list, basing our review on field data issuing from interviews and on our expert knowledge. As a result, we provide a substantially updated list of 292 NWFP growing on the island. Further 34 species, reported as NWFP on previous papers were discarded because they are not native to Sicily, while 45 species were listed separately because their identity, occurrence and local use as food is doubtful and needs to be further investigated. Moreover, we tried to shed light on the ecology (growth form and preferential habitat) of the Sicilian NWFP, with special focus on crop wild relatives (CWR). Our preliminary ecological analyses point out that a high percentage of these plants are linked with the so-called 'cultural' landscapes, patchy semi-natural environments rich in ecotones, leading to the conclusion that the maintenance of century-old agro-pastoral practices may represent an effective way to preserve the local heritage of edible plants. Our study allowed to identify as much as 102 taxa of agronomic interest which could be tested as novel crops in order to face ongoing global changes and to comply with sustainable agriculture policies. Among them, 39 taxa show promising traits in terms of tolerance to one or more environmental stress factors, while 55 more are considered CWR and/or can be easily cultivated and/or show high productivity/yield potential.


Characterization of Tarakeshwara Rasa: An Ayurvedic herbomineral formulation.
Tarakeshwara Rasa (TR) is an Ayurvedic herbomineral compound formulation used in the intervention of Prameha vis-à-vis diabetes mellitus. The present study was executed to establish a fingerprint for this unique formulation, which can be adopted by the Ayurvedic pharmacies for drug standardization. TR is a formulation prepared by the trituration of four ingredients Abhraka Bhasma (AB), Loha Bhasma (LB), Vanga Bhasma (VB) and Rasa Sindhura (RS) in equal quantities with honey for one day. Each of the ingredients were prepared according to the norms of Ayurvedic classical texts and by employing Electric Muffle Furnace as heating device for incineration. To ensure the proper preparation of Bhasmas, standard tests (Bhasma Pariksha) were employed. After Bhasma complies these tests, TR was prepared and subjected for qualitative analysis, X-Ray Diffraction (XRD) and Scanning Electron Microscopy (SEM) studies. LB, AB, VB, RS and TR were also studied for free metal presence by employing phosphomolybdic acid. Chemical analysis of TR reveals that it contains Fe, Sn, Hg, Al, Mn, Ca and Mg. XRD study indicates that TR contains Fe2O3 (maghamite) in major phase and SnO2 (cassiterite), HgS, SiO2, HgO in minor phases. SEM study revealed that the compound is an agglomeration of particles. The particle size was in between 0.5 and 2 μ. Free metal detection by phosphomolybdic acid revealed the absence of free metals in the final Bhasmas. This is the first study establishing the characterization of Tarakeshwara Rasa.


Interplay between Hepatitis E Virus and Host Cell Pattern Recognition Receptors.
Hepatitis E virus (HEV) usually causes self-limiting acute hepatitis, but the disease can become chronic in immunocompromised individuals. HEV infection in pregnant women is reported to cause up to 30% mortality, especially in the third trimester. Additionally, extrahepatic manifestations like neuronal and renal diseases and pancreatitis are also reported during the course of HEV infection. The mechanism of HEV pathogenesis remains poorly understood. Innate immunity is the first line of defense triggered within minutes to hours after the first pathogenic insult. Growing evidence based on reverse genetics systems, in vitro cell culture models, and representative studies in animal models including non-human primates, has implicated the role of the host's innate immune response during HEV infection. HEV persists in presence of interferons (IFNs) plausibly by evading cellular antiviral defense. This review summarizes our current understanding of recognizing HEV-associated molecular patterns by host cell Pattern Recognition Receptors (PRRs) in eliciting innate immune response during HEV infection as well as mechanisms of virus-mediated immune evasion.


Targeting of BMI-1 expression by the novel small molecule PTC596 in mantle cell lymphoma.
Despite the development of the novel Bruton tyrosine kinase inhibitor ibrutinib, mantle cell lymphoma (MCL) remains an incurable B-cell non-Hodgkin lymphoma. BMI-1 is required for the self-renewal and maintenance of MCL-initiating stem cells. Upregulation of BMI-1 has been reported in MCL patients, especially in those with refractory/relapsed disease. We studied the effects of a novel small-molecule selective inhibitor of BMI1 expression, PTC596, in MCL cells. Eight MCL cell lines and patient-derived samples were exposed to PTC596. PTC596 induced mitochondrial apoptosis, as evidenced by loss of mitochondrial membrane potential, caspase-3 cleavage, BAX activation, and phosphatidylserine externalization. There was a positive correlation between baseline BMI-1 protein levels and PTC596-induced apoptosis. p53 status did not affect sensitivity to PTC596. PTC596 effectively decreased BMI-1-expressing and tumor-initiating side population MCL cells (IC50: 138 nM) compared with ibrutinib, which modestly decreased side population cells. Interestingly, PTC596, reported to target cancer stem cells, decreased MCL-1 expression levels and antagonized ibrutinib-induced increase in MCL-1 expression, leading to synergistic apoptosis induction in MCL cells. There are currently no drugs that specifically target cancer stem cell fractions, and a reduction in BMI-1 protein by PTC596 may offer a novel therapeutic strategy for MCL.


Readmission and reoperation after midurethral sling.
We aimed to determine the rate of readmission and reoperation for patients undergoing midurethral sling (MUS) placement for stress urinary incontinence (SUI). The American College of Surgeons National Surgical Quality Improvement Program (NSQIP) database was queried to identify all isolated MUS placed from 2012 through 2015 using the Current Procedural Terminology 4 (CPT-4) code for MUS with or without cystoscopy (57,288 ± 52,000). The cohort was then reviewed for unplanned, related readmissions and reoperations within 30 days of MUS placement. Isolated MUS was placed in 9910 patients. Fifty-eight (0.59%) patients were readmitted and 81 (0.82%) had reoperation. The most common indications for readmission were related to the urinary tract, i.e., urinary retention (27.6%), non-surgical-site-related infection (15.5%), and medical related issues (15.5%) The most common indications for reoperation were urinary tract (60.5%), gastrointestinal (7.4%), and gynecologic, i.e., examination under anesthesia (6.2%). Body mass index (BMI) was less (p = 0.001), and operative time (p = 0.014) and length of stay (LOS) (p = 0.001) longer in patients who were readmitted. Those who underwent reoperation had longer LOS than those who did not have reoperation (p < 0.001). Upon multivariate analysis, BMI <25 (all p < 0.05) and longer LOS maintained statistical significance as risk factors for those who experienced readmission or reoperation (p = 0.0406, p < 0001). Isolated MUS placement has low 30-day readmission and reoperation rates. Increased LOS was associated with readmission, while increased LOS and BMI <25 were associated with reoperation within 30 days.


Egyptian families caring for a relative with mental illness: a hermeneutic study.
The care of people with mental illness often rests with their families. The way families manage this experience has been explored widely in the literature; however, the added complexity of migration has been given little attention. The difficulty of conducting research with people from non-English-speaking backgrounds often precludes their voices and experiences from being heard, further compounding marginalization and misunderstanding of their needs. The aim of this study was to understand the experience of Egyptian families caring for a relative with mental illness in Australia. Seven participants from Egyptian background, caring for a relative with mental illness, were included in this study. A hermeneutic phenomenological approach, informed by the work of Heidegger, was used. Data were collected through in-depth audio-taped interviews conducted in the Arabic language, which were then translated and transcribed in English. Data analysis revealed five themes: Why did it happen? How do I protect my loved ones? What has it done to me? What has it done to us? How do I survive? Findings of the study have the potential to raise health-care professionals' awareness of the needs of Egyptian families, their beliefs, values, and coping with mental illness.


Mechanoreceptor endings in human cervical facet joints.
Twenty-one cervical facet capsules, taken from three normal human subjects, were examined to determine the type, density, and distribution of mechanoreceptive nerve endings in these tissues. Clearly identifiable mechanoreceptors were found in 17 of 21 specimens and were classified according to the scheme for encapsulated nerve endings established by Freeman and Wyke. Eleven Type I, 20 Type II, and 5 Type III receptors were identified, as well as a number of small, unencapsulated nerve endings. Type I receptors were small globular structures measuring 25-50 microns in diameter. Type II receptors varied in size and contour, but were characterized by their oblong shape and broad, lamellated capsule. Type III receptors were relatively large oblong structures with an amorphous capsule, within which a reticular meshwork of fine neurites was embedded. Free (nociceptive) nerve endings were found in subsynovial loose areolar and dense capsular tissues. The presence of mechanoreceptive and nociceptive nerve endings in cervical facet capsules proves that these tissues are monitored by the central nervous system and implies that neural input from the facets is important to proprioception and pain sensation in the cervical spine. Previous studies have suggested that protection muscular reflexes modulated by these types of mechanoreceptors are important in preventing joint instability and degeneration. It is suggested that the surgeon take steps to avoid inadvertently damaging these tissues when exposing the cervical spine.


[Polychemotherapy treatment with cisplatin, adriamycin, 5-fluorouracil (FAP) and cisplatin, adriamycin and cyclophosphamide (CISCA) in advanced transitional carcinoma of the bladder].
We study 50 patients with advanced bladder carcinoma, divided into two protocols of 25 patients, treated with polychemotherapy. Protocol I (PAF) formed by DDP at 20 mgrs/m2 day 1 to 5, ADM at 50 mgrs./m2 day 1 and 5-FU at 500 mgrs./m2 day 1, and Protocol II (CISCA) made up of the combination of DDP at 75 mgrs./m2 day 1, ADM at 50 mgrs./2 day 1 and CPM at 600 mgrs./m2 I.V. day 1. In Protocol I the overall response was 60% (RC = 28%, RP = 32%), with amean response duration of 12.24 months, after receiving an average of 4.8 cycles. In Protocol II the results were 60% (RC = 16%, RP = 44%), 7.04 months and 5 cycles per patient, respectively. Both protocols were tolerated well, although Protcol I proved more toxic. Nauseas, vomiting and alopecia were the most common symptoms. There was no significant difference between the survival of responders and non-responders with Protocol I, but there was with Protocol II with (p less than 0.01).


Carrier testing in hemophilia B with an immunoassay that distinguishes a prevalent factor IX dimorphism.
Immunoassays with a monoclonal antibody (A-1) detect a prevalent dimorphism in plasma coagulation factor IX. The antibody was shown to react with a dimorphic segment of the normal factor IX sequence as follows. First, A-1 bound to isolated activation peptide (residues 146 through 180) prepared from activated factor IX from a normal plasma pool. Second, binding of recombinant factor IXs with A-1 or factor IX from normal individuals was strong only when they had Threonine (Thr) at position 148; factor IXs with the Alanine (Ala) allele at that position were far less reactive. Third, immunoblot reactivity of Escherichia coli fusion proteins containing known segments of the factor IX sequence restricted the epitope to residues 147 through 153. In 120 hemophilia B pedigrees, the Ala immunoassay allele frequency was 0.19 and did not differ from the Ala frequency in normal males. In 22 of 49 families, immunoassay testing was informative for classification of obligate or possible carriers. In one large family, 4 obligate carriers were heterozygous for the dimorphism and 3 of their 7 daughters were classified as carriers. In other families, when the affected member had less than 1 nmol/L factor IX antigen, heterozygosity for Thr/Ala alleles excluded the carrier state even when DNA studies were not informative. Strong linkage disequilibrium of Thr/Ala alleles with the common TaqI DNA polymorphism was found. Nineteen of 75 normal and hemophilic factor IX genes had the 1.3-kilobase (kb) fragment and coded for the Ala allele; the rest had the 1.8-kb fragment and coded for Thr. In selected families, the A-1 immunoassay is an inexpensive and rapid method to confirm and supplement restriction fragment length polymorphism analyses of DNA for carrier testing.


Biomechanical analysis of an aortic aneurysm model and its clinical application to thoracic aortic aneurysms for defining "saccular" aneurysms.
We aimed to develop a simple structural model of aortic aneurysms using computer-assisted drafting (CAD) in order to create a basis of definition for saccular aortic aneurysms. We constructed a simple aortic aneurysm model with 2 components: a tube similar to an aorta and an ellipse analogous to a bulging aneurysm. Three parameters, including the vertical and horizontal diameters of the ellipse and the fillet radius, were altered in the model. Using structural analysis with the finite element method, we visualized the distribution of the maximum principal stress (MPS) in the aortic wall and identified the area(s) of prominent stress. We then selected patients with thoracic aortic aneurysms in whom the aneurysm expansion rates were followed up and applied the theoretical results to the raw imaging data. The maximum MPS drastically increased at areas where the aspect ratio (vertical/horizontal) was <1, indicating that "horizontally long" hypothetical ellipses should be defined as "saccular" aneurysms. The aneurysm expansion rate for the patients with thoracic aneurysms conforming to these parameters was significantly high. Further, "vertically long" ellipses with a small fillet might be candidates for saccular aneurysms; however, the clinical data did not support this. Based on the biomechanical analysis of a simple aneurysm model and the clinical data of the thoracic aortic aneurysms, we defined "horizontally long" aortic aneurysms with an aspect ratio of <1 as "saccular" aneurysms.


Biosynthesis of skin collagens in normal and diabetic mice.
Synthesis of collagens in vitro was studied on minced mouse skins incubated with [3H]-proline in organ-culture conditions. A comparative study was carried out on genetically diabetic mice (KK strain) and control mice (Swiss strain). After incubation, neutral-salt-soluble and acid-soluble collagens were extracted. The insoluble dermis was digested by pepsin and type I and type III collagens separated by differential precipitation in neutral salt solutions. Type I and Type III collagens were characterized by ion-exchange and molecular-sieve chromatography, amino acid analysis and by the characterization of CNBr peptides. In diabetic-mouse skin, the relative proportion of type III collagen was significantly higher than in control-mouse skin. The incorporation of radioactively labelled proline into hydroxyproline of type III collagen was significantly faster in diabetic-mouse skin than in control-mouse skin. No significant modifications in the total collagen content of the skin or of their rates of synthesis were observed between the two strains. Alteration in the ratio of type III to type I collagen in the diabetic-mouse skin can be interpreted as a sign of alteration of the regulation of collagen biosynthesis and may be related to the structural alterations observed in the diabetic intercellular matrix.


Stress measurement using speech: Recent advancements, validation issues, and ethical and privacy considerations.
Life stress is a well-established risk factor for a variety of mental and physical health problems, including anxiety disorders, depression, chronic pain, heart disease, asthma, autoimmune diseases, and neurodegenerative disorders. The purpose of this article is to describe emerging approaches for assessing stress using speech, which we do by reviewing the methodological advantages of these digital health tools, and the validation, ethical, and privacy issues raised by these technologies. As we describe, it is now possible to assess stress via the speech signal using smartphones and smart speakers that employ software programs and artificial intelligence to analyze several features of speech and speech acoustics, including pitch, jitter, energy, rate, and length and number of pauses. Because these digital devices are ubiquitous, we can now assess individuals' stress levels in real time in almost any natural environment in which people speak. These technologies thus have great potential for advancing digital health initiatives that involve continuously monitoring changes in psychosocial functioning and disease risk over time. However, speech-based indices of stress have yet to be well-validated against stress biomarkers (e.g., cortisol, cytokines) that predict disease risk. In addition, acquiring speech samples raises the possibility that conversations intended to be private could one day be made public; moreover, obtaining real-time psychosocial risk information prompts ethical questions regarding how these data should be used for medical, commercial, and personal purposes. Although assessing stress using speech thus has enormous potential, there are critical validation, privacy, and ethical issues that must be addressed.


Importance of Video-EEG Monitoring in the Diagnosis of Epilepsy in a Psychiatric Patient.
Epilepsy is a chronic medical condition which is disabling to both patients and caregivers. The differential diagnosis of epilepsy includes psychogenic nonepileptic spells or "pseudoseizures." Epilepsy is due to abnormal electrical activity in the brain, and pseudoseizure is a form of conversion disorder. The brain waves remain normal in pseudoseizures. The problem arises when a patient with significant psychiatric history presents with seizures. Pseudoseizures become high on the differential diagnosis without extensive work up. This is a case of woman with significant psychiatric issues which resulted in a delay in the diagnosis of epilepsy.


Optimizing the treatment of ovarian cancer: Neoadjuvant chemotherapy and interval debulking versus primary debulking surgery for epithelial ovarian cancers likely to have suboptimal resection.
To: a) identify prognostic factors in patients with epithelial ovarian cancer treated with neoadjuvant chemotherapy (NACT) and interval debulking surgery (IDS), and b) compare post-surgical survival between patients treated with NACT/IDS for presumed unresectable disease and stage IIIC/IV patients who underwent suboptimal primary debulking surgery (PDS). This was a retrospective study of consecutive stage IIIC or IV patients undergoing IDS after NACT at Mayo Clinic from January 2007 to December 2013. A subset of patients receiving NACT/IDS for the indication of unresectable disease were matched 1:1 on age and stage to a cohort of patients who underwent suboptimal PDS between 2003 and 2011. Hazard ratios and corresponding 95% confidence intervals were estimated from Cox proportional hazards models. We identified 87 patients treated with NACT/IDS: the median OS and PFS following surgery was 2.4 and 1.0years, respectively. Factors associated with significantly worse OS were older age (adjusted HR 1.60 per 10-year increase in age, 95% CI 1.18, 2.16) and elevated CA-125 before IDS (adjusted HR 2.30 for CA-125 >35U/mL, 95% CI 1.25, 4.23). Number of adjuvant chemotherapy cycles administered did not have a significant effect on survival. In the matched cohort analysis of presumed unresectable cases undergoing NACT/IDS vs suboptimal PDS cases (n=45 each), the NACT/IDS group had a significant OS advantage (HR 0.53; 95% CI 0.32, 0.88), and fewer patients experienced a 30-day postoperative Accordion grade 3/4 complication (11% vs 36%, P=0.01). Younger age and normalization of CA-125 prior to IDS are associated with improved survival with NACT/IDS. For primary EOC where resection to residual disease of 1cm or less is unlikely, NACT/IDS is associated with improved survival and reduced perioperative morbidity compared to PDS. As these patients are likely best served by NACT/IDS, more reliable predictors of resectability would be valuable.


Brain benzodiazepine receptors in fathead minnows and the behavioral response to alarm pheromone.
Fathead minnows (Pimephales promelas) exposed to 1 or 10 mg/l chlordiazepoxide showed normal alarm behavior during the presentation of alarm pheromone. Fish exposed to 20 mg/l drug, however, showed little or no behavioral alarm and did not appear sedated. A food extract stimulus presented after alarm pheromone led to a large foraging response by fish exposed to 20 mg/l chlordiazepoxide. Control fish or fish exposed to 1 to 10 mg/l drug showed less tendency to begin foraging. Exposure to 1, 10, or 20 mg/l chlordiazepoxide for 3 hr did not affect whole-brain concentrations of tryptophan, 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, tyrosine, or dopamine. The binding of [3H]diazepam to specific brain receptor sites (KD = 10 nM, estimated Bmax = 3.3 fmol/mg wet weight) could be displaced by chlordiazepoxide (IC50 = 1.6 microM). These results suggest that benzodiazepine receptors in the central nervous system of lower vertebrates may function in ways similar to those in mammals, i.e., in the modulation of behavior in anxiety-like states.


Mutational analysis of the propensity for amyloid formation by a globular protein.
Acylphosphatase can be converted in vitro, by addition of trifluoroethanol (TFE), into amyloid fibrils of the type observed in a range of human diseases. The propensity to form fibrils has been investigated for a series of mutants of acylphosphatase by monitoring the range of TFE concentrations that result in aggregation. We have found that the tendency to aggregate correlates inversely with the conformational stability of the native state of the protein in the different mutants. In accord with this, the most strongly destabilized acylphosphatase variant forms amyloid fibrils in aqueous solution in the absence of TFE. These results show that the aggregation process that leads to amyloid deposition takes place from an ensemble of denatured conformations under conditions in which non-covalent interactions are still favoured. These results support the hypothesis that the stability of the native state of globular proteins is a major factor preventing the in vivo conversion of natural proteins into amyloid fibrils under non-pathological conditions. They also suggest that stabilizing the native states of amyloidogenic proteins could aid prevention of amyloidotic diseases.


[The future of laparoscopic surgery in urology].
Laparoscopy has followed the typical phases in the development of any new operative technique. We have now begun to realize its limitation and are beginning to standardize the procedures and specify their indications. Since laparoscopy predominantly reduces the trauma of access, the development of a total operative trauma score has assisted us in determining the impact of specific laparoscopic procedures compared to their open alternatives. Laparoscopy is responsible for less blood loss, less postoperative pain, a shorter hospital stay and a very small scar, all of which accounts for its popularity among patients. A rational combination of laparoscopy, endoscopy and open surgery to treat urological diseases should be developed in the future.


Prevention and treatment of Staphylococcus biofilms.
Staphylococcus growth on medical devices represents a common occurrence that can lead to serious illness and death. Biomaterial-associated infection, mostly caused by Staphylococcus epidermidis and Staphylococcus aureus, is fairly complicated by the organism' development of a biofilm, which provides a microenvironment that protects from attack by the host immune system and antibiotics. In this review we present recent insights regarding S. aureus and S. epidermidis structural and functional factors that are effective in biofilm development and describe the regulation of their expression. On the basis of the knowledge gained, we also present the potential and limits of current biochemical and biophysical strategies aimed at preventing biofilm formation or at the treatment of established mature biofilms.


[Respiratory gated radiotherapy: the 4D radiotherapy].
Respiration-gated radiotherapy offers a significant potential for improvement in the irradiation of tumor sites affected by respiratory motion such as lung, breast and liver tumors. An increased conformality of irradiation fields leading to decreased complications rates of organs at risk (lung, heart...) is expected. Respiratory gating is in line with the need for improved precision required by radiotherapy techniques such as 3D conformal radiotherapy or intensity modulated radiotherapy. Reduction of respiratory motion can be achieved by using either breath hold techniques or respiration synchronized gating techniques. Breath hold techniques can be achieved with active, in which airflow of the patient is temporarily blocked by a valve, or passive techniques, in which the patient voluntarily breath-hold. Synchronized gating techniques use external devices (CCD camera for the GEMS/Varian system tested at Curie Institute) to predict the phase of the respiration cycle while the patient breaths freely. A new strategy is currently developed: the 4D Respiration correlated CT. It consists of retrospectively reconstruct CT slices at different phases of the breathing cycle allowing to measure residual movements and to choose the optimal patient's breathing phase where tumor movements are lower. These techniques presently investigated in several medical centers worldwide. The first results are very promising.


Ultrastructural surface alterations of guinea pig seminal vesicle epithelium during postnatal development and after castration.
We have designed a topographic anatomic study using scanning electron microscopy to define surface changes in seminal vesicle epithelium during normal development and after bilateral orchiectomy. Scanning electron microscopy of seminal vesicles from 1-day, 4-week, and 3-month-old intact animals revealed a microvilli-studded luminal epithelial surface with high polyhedral cell profiles extending into the luminal area. Deep furrows outlined all intercellular junctions. Microvilli profiles ranged from short knob-like projections located preferentially along cell margins in 1-day-old animals to 1 mu-long microvilli wih random distribution in mature animals. The most striking finding was that the seminal vesicle epithelium in castrated animals did not revert to the prepubertal state. Rather, cell apices receded from the luminal surface to a level below that of the cell borders and microvilli generally remained somewhat longer in the castrate than in the prepubertal animal. These morphologic findings suggest that the seminal vesicle epithelium in the prepubertal animal and the long-term castrate are quite different.


Resistance to murine cytomegalovirus linked to the major histocompatibility complex of the mouse.
Studies of the resistance patterns to infection with a murine cytomegalovirus in inbred strains of mice revealed the existence of resistant and susceptible strains. Resistance was found to be associated with possession of the H-2k allele at the major histocompatibility locus of the mouse. The F1 hybrid between a resistant strain (C3H/HeJ) and a susceptible strain (BALB/c) was found to have a resistance intermediate between that of both parents, indicating that the gene(s) controlling resistance is partly dominant. Susceptible BALB/c mice could be made resistant to lethal infection by pre-treatment with thioglycollate broth but not by pre-treatment with endotoxin or BCG. Resistant C3H/HeJ mice could be made susceptible to lethal infection by pre-treatment with cyclophosphamide.


Successful laparoscopic splenorrhaphy using absorbable mesh for grade III splenic injury: report of a case.
We report herein on successfully performing a laparoscopic mesh splenorrhaphy in addition to blood salvage on a patient with a grade III splenic injury from blunt abdominal trauma. Given the lower success rates for nonoperative observation in grade III splenic trauma, we believe laparoscopic evaluation of splenic injury may offer an improved assessment over computed tomography (CT) scanning by localizing and quantitating the source of blood loss while at the same time enabling the surgeon to perform laparoscopically assisted blood salvage, autotransfusion, and laparoscopic repair. These advantages in diagnosis and therapy warrant further investigation into the role of laparoscopy in the evaluation and treatment of splenic trauma in selected stable patients.


Fetal programming of body composition, obesity, and metabolic function: the role of intrauterine stress and stress biology.
Epidemiological, clinical, physiological, cellular, and molecular evidence suggests that the origins of obesity and metabolic dysfunction can be traced back to intrauterine life and supports an important role for maternal nutrition prior to and during gestation in fetal programming. The elucidation of underlying mechanisms is an area of interest and intense investigation. In this perspectives paper we propose that in addition to maternal nutrition-related processes it may be important to concurrently consider the potential role of intrauterine stress and stress biology. We frame our arguments in the larger context of an evolutionary-developmental perspective that supports roles for both nutrition and stress as key environmental conditions driving natural selection and developmental plasticity. We suggest that intrauterine stress exposure may interact with the nutritional milieu, and that stress biology may represent an underlying mechanism mediating the effects of diverse intrauterine perturbations, including but not limited to maternal nutritional insults (undernutrition and overnutrition), on brain and peripheral targets of programming of body composition, energy balance homeostasis, and metabolic function. We discuss putative maternal-placental-fetal endocrine and immune/inflammatory candidate mechanisms that may underlie the long-term effects of intrauterine stress. We conclude with a commentary of the implications for future research and clinical practice.


Intracellular staining of Mx proteins in cells from peripheral blood, bone marrow and skin.
The Mx proteins are known to be specifically and dose dependently induced in mononuclear cells (MNC) by type I interferons (IFN). The aim of this study was to establish a staining method for the human intracellular Mx proteins, MxA and MxB, in leucocytes and bone marrow and skin cells. Several monoclonal antibodies directed against the MxA and MxB proteins were generated. These antibodies were used to stain Mx proteins in both frozen and paraffin wax sections using the standard alkaline phosphatase anti-alkaline phosphatase (APAAP) method. Granulocytes, monocytes and lymphocytes extracted from freshly collected blood from 21 healthy subjects did not stain. After incubating MNC from these subjects with IFN alpha 2b for 48 hours, Mx proteins were detected in monocytes and lymphocytes. Within two days of starting treatment with subcutaneous IFN alpha 2b, granulocytes, monocytes and lymphocytes of 16 patients with cancer stained strongly for Mx proteins. The intensity of staining was correlated with the Mx content of whole blood measured using a specific ELISA. Prior to IFN treatment, cells from bone marrow and skin tissue specimens were negative for Mx proteins with the exception of endothelial cells. During treatment with IFN alpha 2b, nearly all cells from bone marrow and skin stained intensely. These new monoclonal antibodies facilitate the detection of Mx positive cells in peripheral blood and in frozen or paraffin wax specimens. The advantage of this staining method is that individual cells which have responded to viruses or biologically active IFN alpha, beta or omega can be identified.


Interaction pattern of Arg 62 in the A-pocket of differentially disease-associated HLA-B27 subtypes suggests distinct TCR binding modes.
The single amino acid replacement Asp116His distinguishes the two subtypes HLA-B*2705 and HLA-B*2709 which are, respectively, associated and non-associated with Ankylosing Spondylitis, an autoimmune chronic inflammatory disease. The reason for this differential association is so far poorly understood and might be related to subtype-specific HLA:peptide conformations as well as to subtype/peptide-dependent dynamical properties on the nanoscale. Here, we combine functional experiments with extensive molecular dynamics simulations to investigate the molecular dynamics and function of the conserved Arg62 of the α1-helix for both B27 subtypes in complex with the self-peptides pVIPR (RRKWRRWHL) and TIS (RRLPIFSRL), and the viral peptides pLMP2 (RRRWRRLTV) and NPflu (SRYWAIRTR). Simulations of HLA:peptide systems suggest that peptide-stabilizing interactions of the Arg62 residue observed in crystal structures are metastable for both B27 subtypes under physiological conditions, rendering this arginine solvent-exposed and, probably, a key residue for TCR interaction more than peptide-binding. This view is supported by functional experiments with conservative (R62K) and non-conservative (R62A) B*2705 and B*2709 mutants that showed an overall reduction in their capability to present peptides to CD8+ T cells. Moreover, major subtype-dependent differences in the peptide recognition suggest distinct TCR binding modes for the B*2705 versus the B*2709 subtype.