Prospective follow-up of female sexual function after vaginal surgery for pelvic organ prolapse using transobturator mesh implants.
Although the use of transobturator mesh implants for pelvic organ prolapse repair has been shown to be safe and effective, concern exists that the presence of prosthetic material in the vagina may adversely affect sexual function. To evaluate the impact of transobturator mesh implantation on sexual function using validated questionnaire. Female Sexual Function Index (FSFI), a validated 19-item questionnaire that assesses six domains of sexual function (desire, arousal, lubrication, orgasm, satisfaction, and pain), was used. The questionnaire was administered preoperatively, and at 3, 6, 12, and 24 months postoperatively. Clinical data were also recorded at each time point. Prospective nonrandomized study including 96 women with pelvic organ prolapse (cystocele, rectocele, vault prolapse). Transvaginal anterior or posterior wall repair using transobturator mesh implants with or without concomitant transobturator sling procedure. Mean age was 51.4 ± 5.2 years. Mean operating time was 47.6 ± 23.4 minutes, and the mean hospitalization period was 3.8 ± 1.6 days. After initial decrease during the first 3 months, patients experienced a steady improvement in their sexual function. At 24 months postoperatively, the total mean FSFI score reached significantly higher values compared to the baseline (P = 0.023). Furthermore, pain-free intercourse improved during the follow-up reaching mean score of 4.27 ± 0.79 (P < 0.05) after 2 years. Pelvic floor examination at 2 years follow-up showed excellent surgical results with only 3.1% of the patients presenting with stage II vaginal wall prolapse. Surgical repair of symptomatic pelvic organ prolapse using mesh implants results in improvement of major parameters of sexual function. A worsening in pain with intercourse during the initial months postoperatively lessens after 3 months as healing is completed.


Donor symptomatology and safety in blood cell separation procedures.
Increasing demands for platelet and granulocyte concentrates collected by cell separation technology for replacement strategies requires expanding available donor pools, related or unrelated by recruiting additional donors or increasing the frequency with which a donor is processed. The expanding application of cell separation devices for plasma or red cell exchange as a therapeutic modality for many medical diagnoses now exposes patients with altered physiology to technology and procedures originally designed for healthy blood component donors. Some possible risks from the technology and/or procedure are discussed. Donor symptomatology from the chemical agents used in blood processing and a relationship to scheduling frequency are introduced.


Discovery of adamantyl ethanone derivatives as potent 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) inhibitors.
11Beta-hydroxysteroid dehydrogenases (11beta-HSDs) are key enzymes regulating the pre-receptor metabolism of glucocorticoid hormones. The modulation of 11beta-HSD type 1 activity with selective inhibitors has beneficial effects on various conditions including insulin resistance, dyslipidemia and obesity. Inhibition of tissue-specific glucocorticoid action by regulating 11beta-HSD1 constitutes a promising treatment for metabolic and cardiovascular diseases. A series of novel adamantyl ethanone compounds was identified as potent inhibitors of human 11beta-HSD1. The most active compounds identified (52, 62, 72, 92, 103 and 104) display potent inhibition of 11beta-HSD1 with IC(50) values in the 50-70 nM range. Compound 72 also proved to be metabolically stable when incubated with human liver microsomes. Furthermore, compound 72 showed very weak inhibitory activity for human cytochrome P450 enzymes and is therefore a candidate for in vivo studies. Comparison of the publicly available X-ray crystal structures of human 11beta-HSD1 led to docking studies of the potent compounds, revealing how these molecules may interact with the enzyme and cofactor.


ALS-Related Mutant FUS Protein Is Mislocalized to Cytoplasm and Is Recruited into Stress Granules of Fibroblasts from Asymptomatic FUS P525L Mutation Carriers.
Amyotrophic lateral sclerosis (ALS) shows a strong genetic basis, with SOD1, FUS, TARDBP, and C9ORF72 being the genes most frequently involved. This has allowed identification of asymptomatic mutation carriers, which may be of help in understanding the molecular changes preceding disease onset. We studied the cellular expression of FUS protein and the effect of heat-shock- and dithiothreitol-induced stress in fibroblasts from FUS P525L mutation carriers, healthy controls, and patients with sporadic ALS. Western blots and immunocytochemistry were performed to study the subcellular localization of FUS protein. Control and stressed cells were double stained with FUS and the stress marker TIA-R. Fibroblasts from healthy controls and sporadic ALS cases showed a prominent nuclear FUS expression. In the 2 FUS P525L mutation carriers, instead, most cells showed a protein localization in both nucleus and cytoplasm, or exclusively in the cytoplasm. Stress prompted the formation of cytoplasmic granules in all subjects and in sporadic ALS FUS mislocalization to the cytoplasm. Cytoplasmic FUS was recruited into stress granules, which showed a time-dependent decrease in all subjects. However, in the FUS P525L fibroblasts, the granules persisted longer, and they were more numerous than those detected in the cells from controls and sporadic ALS patients. We show that in fibroblasts of FUS P525L mutation carriers, FUS mislocalized to the cytoplasm where it redistributed into stress granules with likely a dose effect, i.e. a higher number of cells with granules, which persist longer, than in controls and ALS cases. These data represent an early molecular change occurring before ALS onset, suggesting a transient preaggregative state.


[Increased CK activity in serum without symptoms: further investigations often unnecessary].
Increased activity of the enzyme creatine kinase (CK) in serum is not infrequently encountered in routine diagnostic laboratory investigations. Patients are often referred to a neurologist specialized in neuromuscular disorders for evaluation. However, as in many cases hyperCKemia is physiological or results from physical activity or muscle trauma, further investigations are often unnecessary. We report four cases of hyperCKemia, two of which were physiological or due to non-neuromuscular factors (medication, physical activity). In the other two patients, the hyperCKemia was the first recognized sign of an underlying neuromuscular disorder. In these two cases, specific aspects of the history or physical examination prompted further investigations. We discuss various physiological and other non-neuromuscular factors that may cause hyperCKemia. It is important to recognize these causes before referral to a neurologist with neuromuscular expertise. We present guidelines for ancillary investigations by general practitioners or specialists.


Sub-national assessment of inequality trends in neonatal and child mortality in Brazil.
Brazil's large socioeconomic inequalities together with the increase in neonatal mortality jeopardize the MDG-4 child mortality target by 2015. We measured inequality trends in neonatal and under five mortality across municipalities characterized by their socio-economic status in a period where major pro poor policies were implemented in Brazil to infer whether policies and interventions in newborn and child health have been successful in reaching the poor as well as the better off. Using data from the 5,507 municipalities in 1991 and 2000, we developed accurate estimates of neonatal mortality at municipality level and used these data to investigate inequality trends in neonatal and under five mortality across municipalities characterized by socio-economic status. Child health policies and interventions have been more effective in reaching the better off than the worst off. Reduction of under five mortality at national level has been achieved by reducing the level of under five mortality among the better off. Poor municipalities suffer from worse newborn and child health than richer municipalities and the poor/rich gaps have increased. Our analysis highlights the importance of monitoring progress on MDGs at sub-national level and measuring inequality gaps to accurately target health and inter-sectoral policies. Further efforts are required to improve the measurement and monitoring of trends in neonatal and under five mortality at sub-national level, particularly in developing countries and countries with large socioeconomic inequalities.


An in vitro and in vivo characterization of fine WE43B magnesium wire with varied thermomechanical processing conditions.
Absorbable implants made of magnesium alloys may revolutionize surgical intervention, and fine magnesium wire will be critical to many applications. Functionally, the wires must have sufficient mechanical properties to withstand implantation and in-service loading, have excellent tissue tolerance, and exhibit an appropriate degradation rate for the application. Alloy chemistry and thermomechanical processing conditions will significantly impact the material's functional performance, but the exact translation of these parameters to implant performance is unclear. With this in mind, fine (127 µm) WE43B magnesium alloy wires in five thermomechanical process (TMP) conditions (90% cold work [CW], and 250, 375, 400, and 450°C heat treatments) were investigated for their effect on mechanical and corrosion behavior. The TMP conditions gave clear metallurgical differences: transverse grain dimensions ranged from 200 nm (CW) to 3 µm (450°C), UTS varied from 324 MPa (450°C) to 608 MPa (250°C), and surgical knotting showed some were suitable (CW, 400°C, 450°C) while others were not (250°C, 350°C). In vitro and in vivo corrosion testing yielded interesting and in some cases conflicting results. After 1 month immersion in cell culture medium, wire corrosion was extensive, and TMP conditions altered the macrocorrosion morphology but not the rate or total release of magnesium ions. After 1 month subdermal implantation in mice, all wires were well tolerated and showed very little corrosion (per µCT and histology), but differences in localized corrosion were detected between conditions. This study indicates that WE43B wires treated at 450°C may be most suitable for surgical knotting procedures. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1987-1997, 2018.


Primary care experience and racial disparities in self-reported health status.
Access to high quality primary care was identified by Healthy People 2010 as one of the mechanisms through which racial and ethnic disparities in health might be reduced. Despite the well-established connections between good primary care and health, the scientific evidence on whether good primary care can reduce racial disparities in health is sparse. To examine whether better primary care experience can attenuate racial and ethnic disparities in self-reported health status. The 1996 to 1997 and 1998 to 1999 data from the Community Tracking Study (CTS) sponsored by the Robert Wood Johnson Foundation. Cross-sectional, bivariate, and multivariate analyses of inter-relationships between self-rated general and mental health status, access to and interpersonal relationship with primary care provider, and vulnerability measured by race and poverty status. We found that higher quality primary care levels are associated with reduced racial and ethnic disparities in health status, as measured by self-rated general and mental health. This relationship is particularly pronounced for the racial and ethnic minorities living at or below poverty level. Based on the data from 1996 to 1999, the study also confirmed the presence of significant and persistent health differences across racial and ethnic groups. Promoting primary care may be a viable approach toward reducing racial and ethnic disparities in self-reported health status.


Central obesity and breast cancer risk: a systematic review.
The specific effect of central rather than general obesity on breast cancer risk is not clear. This review examines the relationship between waist and waist-hip ratio (WHR) and risk of breast cancer in pre- and post-menopausal women using all available cohort and case-control data. The databases of the Cochrane Library, Medline, Cancer Lit and Embase were searched until October 2002. Relevant cohort and case-control studies with separate analyses in pre- and/or post-menopausal women were included. Random effects meta-analyses were carried out, subgrouped by pre- or post-menopausal status and cohort or case-control design. Sensitivity analyses were also performed. Five cohort studies with 72,1705 person years of observation (453 pre-menopausal and 2684 post-menopausal cases), and three case-control studies comprising 276 pre-menopausal cases with 758 pre-menopausal controls and 390 post-menopausal cases with 1071 post-menopausal controls were included. Pooled results from cohort studies using the most adjusted data [but without adjustment for weight or body mass index (BMI)] suggest a 39% lower risk of breast cancer in post-menopausal women with the smallest waist (compared with the largest) and a 24% lower risk in women with the smallest WHR. In pre-menopausal women, however, pooled results suggest that measurement of waist or WHR have little effect on risk of breast cancer. Adjustment for BMI abolished the relationship between waist or WHR and risk of post-menopausal breast cancer, but introduced such a relationship amongst pre-menopausal women. The relationship between a smaller measurement of waist or WHR and lower risk of post-menopausal breast cancer appears to result from the associated correlation with BMI. Amongst pre-menopausal women, central (not general) obesity may be specifically associated with an increased risk of breast cancer.


[Timing of surgical intervention in acute spinal cord injury and post-operative neurological recovery].
The effect of an early surgical intervention in the traumatised spine on resolution of neurological deficit still remains a topic of professional discussions. The aim of this retrospective study was to find a correlation between the length of an injury-to-surgery interval and the development of a post-operative neurological deficit, and thus to answer the question of whether early surgical decompression and stabilization gives better chance of neurological recovery. Medical records of consecutive surgical patients admitted between 2007 and 2010 with traumatic spinal cord injury were reviewed and the injury-to-surgery interval and post-operative development of neurological deficit at a minimum follow-up of 6 months was evaluated. The initial neurological finding and the finding at 6 months of follow-up were classified on the Frankel scale and the outcome was assessed as improved or unimproved. The patients were allocated to four subgroups according to the time that elapsed between injury and surgery, i.e., time up to 24 h, 24-72 h, 72 h -1 week, and longer than 1 week. The percentage of improved patients was calculated in each subgroup and the results were statistically evaluated using the Kruskal-Wallis test at a significance level of 0.1. Out of the total number of 32 evaluated patients, 28 had at least partial neurological recovery. In the subgroup treated within first 24 h, improvement was found in 93 % of the patients, in the 24-72 h subgroup it was 80%, in the 72 h-1 week subgroup it was 60% and surgery later than a week after injury resulted in improvement in 42% of the patients. Based on statistical evaluation, the time between injury and surgery appeared to be a significant prognostic factor. When a paired comparison of subgroups was made, the only significant difference was found between the subgroup treated within 24 hours of injury and that operated on later than a week after injury. The other paired comparisons failed to show a significant difference due to a small number of patients; however, a tendency to better functional results was observed in all earlier- treated subgroups. The authors are aware of few limitations of the study. Its retrospective character, a relatively small number of patients and a single institution setup may limit the interpretation. Despite this fact, the message is clear. Similar studies carried out prospectively at several institutions may, however, provide results with a higher validity. Patients with traumatic spinal cord injury who undergo early decompression and stabilisation have a higher chance of at least partial neurological recovery.


COVID-19 spreading in Rio de Janeiro, Brazil: Do the policies of social isolation really work?
The recent Coronavirus has been spreading through all the world fastly. In this work we focus on the evolution of the COVID-19 in one of the most populous Brazilian states, namely the Rio de Janeiro state. The first case was reported in March 5, 2020, thus we have a considerable amount of available data to make a good analysis. We study the dynamics of COVID-19 through a Susceptible-Infectious-Quarantined-Recovered (SIQR) model with an additional mechanism that represents the implementation of public policies of social isolation. Based on the data collected from the Rio de Janeiro state Department of Health from March 5, 2020 through April 26, 2020, we observed that the implementation of social distancing policies changed the initial exponential growth to a sub-exponential one. The SIQR model with the above-mentioned mechanism is capable of reproducing the data of confirmed cases in Rio de Janeiro, and it explains how that change occurred. The model also predicts an important mitigation effect, namely the flattening effect, i.e., the considerably decrease of the maximum number of confirmed cases. Through the results of the model, this effect can be directly related to the social isolation policies. Finally, we consider the relaxation of such policies, and discuss about the ideal period of time to release people to return to their activities.


The application of genetically engineered herpes simplex viruses to the treatment of experimental brain tumors.
Due to lack of effective therapy, primary brain tumors are the focus of intense investigation of novel experimental approaches that use vectors and recombinant viruses. Therapeutic approaches have been both indirect, whereby vectors are used, or direct to allow for direct cell killing by the introduced virus. Genetically engineered herpes simplex viruses are currently being evaluated as an experimental approach to eradicate malignant human gliomas. Initial studies with gamma (1)34.5 mutants, R3616 (from which both copies of the gamma (1)34.5 gene have been deleted) and R4009 (a construct with two stop codons inserted into the gamma (1)34.5 gene), have been assessed. In a syngeneic scid mouse intracranial tumor model, recombinant herpes simplex virus can be experimentally used for the treatment of brain tumors. These viruses and additional engineered viruses were subsequently tested in human glioma cells both in vitro and in vivo. Using a xenogeneic scid mouse intracranial glioma model, R4009 therapy of established tumors significantly prolonged survival. Most importantly, long-term survival was achieved, with histologic evidence that R4009 eradicated intracranial tumors in this model. Furthermore, the opportunity to evaluate gamma (1)34.5 mutants that have enhanced oncolytic activity, e.g., R8309 where the carboxyl terminus of the gamma (1)34.5 gene has been replaced by the murine homologue, MyD116, are considered.


Compassion Practices, Nurse Well-Being, and Ambulatory Patient Experience Ratings.
Compassion practices both recognize and reward compassion in the workplace as well as provide compassionate support to health care employees. However, these practices represent an underexplored organizational tool that may aid clinician well-being and positively impact patient ambulatory care experiences. To examine the relationship between compassion practices and nursing staff well-being and clinic-level patients' experience ratings in the ambulatory clinic setting. Surveys were collected from ambulatory nurses in January and February of 2015 in 30 ambulatory clinics affiliated with an academic medical center. Patient experience ratings were collected April to June of 2015. One hundred seventy-seven ambulatory nurses (Registered Nurses, LPNs, medical assistants), as well as 3525 adult patients from the ambulatory clinics. Ambulatory nurses assessed compassion practices, emotional exhaustion, and psychological vitality. Patient experience ratings were patient perceptions of courtesy and caring shown by nurses and patients' ratings of the outpatient services. Compassion practices are significantly and negatively associated with nurse emotional exhaustion and positively associated with nurse psychological vitality. At the clinic-level, compassion practices are significantly and positively associated with patient perceptions of caring shown by nurses and overall patient ratings of the outpatient clinic. Supplemental analyses provide preliminary evidence that nurse well-being mediates the relationship between compassion practices and patient ratings of their care experience. Our findings illustrate that compassion practices are positively associated with nurse well-being and patient perceptions of the care experience in outpatient clinics.


Determining the activity of mucosal adjuvants.
Mucosal vaccination offers the advantage of blocking pathogens at the portal of entry, improving patient's compliance, facilitating vaccine delivery, and decreasing the risk of unwanted spread of infectious agents via contaminated syringes.Recent advances in vaccinology have created an array of vaccine constructs that can be delivered to mucosal surfaces of the respiratory, gastrointestinal, and genitourinary tracts using intranasal, oral, and vaginal routes. Due to the different characteristics of mucosal immune response, as compared with systemic response, mucosal immunization requires particular methods of antigen presentation. Well-tolerated adjuvants that enhance the efficacy of such vaccines will play an important role in mucosal immunization. Among promising mucosal adjuvants, mutants of cholera toxin and the closely related heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli present powerful tools, augmenting the local and systemic serum antibody response to co-administered antigens.In this chapter, we describe the formulation and application of vaccines using the genetically modified LTK63 mutant as a prototype of the family of these mucosal adjuvants and the tools to determine its activity in the mouse model.


Exogenous abscisic acid reduces water loss and improves antioxidant defence, desiccation tolerance and transpiration efficiency in two spring wheat cultivars subjected to a soil water deficit.
The effect of soil drenching with 10µM abscisic acid (ABA) on the physiological responses of two spring wheat (Triticum aestivum L.) cultivars released in different decades was evaluated when subjected to a water deficit at jointing or at booting. Exogenous ABA application increased the ABA concentration in the leaves, reduced the stomatal conductance (gs), slowed the rate of water use, decreased the lethal leaf water potential (ψ) used to measure desiccation tolerance and lowered the soil water content (SWC) at which leaf relative water content (RWC) began to decrease and wilting was observed. Exogenous ABA application also reduced reactive oxygen species (ROS) formation and increased antioxidant enzyme activity, leading to a reduction in the oxidative damage to lipid membranes in both cultivars exposed to water stress at jointing and booting. The decrease in leaf RWC and wilting occurred at lower values of SWC in the recently-released cultivar than in the earlier-released cultivar. The recently-released cultivar also had higher grain yield than the earlier-released cultivar at moderate water stress, but the grain yield in both cultivars was reduced by water stress and by the exogenous ABA treatment. However, exogenous ABA treatment increased transpiration efficiency for grain (TEG) of both cultivars under moderate water stress. These results indicate that ABA played an important role in slowing water use and enhancing the antioxidant defence during soil drying, but this did not result in increased yields under drought stress.


Safety and efficacy of pressure-limited power injection of iodinated contrast medium through central lines in children.
The purpose of this study was to evaluate the safety and efficacy of pressure-limited power injection of contrast medium through central lines for pediatric body CT examinations. All patients with a central line who were referred for body CT examinations requiring an i.v. contrast agent were prospectively evaluated. The power injector was pressure limited to 25 psi (172 kPa). A standard dose of 2 mL/kg of iodinated contrast medium was power-injected through the central line. Two pediatric radiologists scored all examinations on a scale of 1 (poor) to 5 (superior) for adequacy of contrast enhancement. Regression and receiver operating characteristic analyses were performed. The subjects were 63 patients 0.3-22 years old. Nineteen of these patients had tunneled lines, 18 had ports, and 26 had peripherally inserted central catheters. There were no complications related to power injection. Regression analysis showed a significant association between patient weight and contrast enhancement adequacy score (p < 0.001), higher patient weights yielding lower contrast enhancement adequacy scores. Receiver operating characteristic analysis showed a weight cutoff of 30 kg as a reasonable predictor of adequacy of contrast enhancement. For patients weighing 30 kg or more, the average contrast enhancement score was 2.4 (suboptimal to adequate). For patients weighing less than 30 kg, the average contrast enhancement score was 3.4 (adequate to good). Pressure-limited power injection through central lines in children is safe. The contrast enhancement obtained with 25 psi (172 kPa) pressure-limited injection is acceptable only for patients who weigh less than 30 kg.


Evaluation of the optimal laparoscopic method for benign ovarian mass extraction: a transumbilical route using a bag made from a surgical glove versus a lateral transabdominal route employing a standard endobag.
We compared two transumbilical (TU) routes of surgical specimen retrieval in women with ovarian masses treated via laparoscopy: a bag made from a surgical glove and lateral transabdominal (LTA) retrieval employing a standard endobag. A total of 109 women undergoing laparoscopic surgery to treat benign adnexal masses were retrospectively evaluated between 2014 and 2017. In total, 57 masses were removed via the TU route and 52 via the LTA route. We recorded the ovarian mass size; additional postoperative analgesic drug requirements. Postoperative incisional pain scores were assessed using a 10-cm visual analogue scale (VAS), time to discharge and procedure type. The mean VAS scores at 1 h (5.0 ± 1.7 vs. 6.3 ± 1.3; p < .001); 12 h (0.7 ± 0.8 vs. 1.2 ± 1.1; p = .004); and 24 h (0.1 ± 0.3 vs. 0.7 ± 0.6; p < .001) were lower in the TU-removal group. Furthermore, additional postoperative analgesic drug requirements were significantly higher in the LTA-removal group (10 (19.2%) vs. 3 (5.3%); p = .03). During laparoscopic surgery, removal of an ovarian mass via an umbilical port (compared to a lateral port) causes less postoperative pain and does not increase the risk of wound complications such as infection or hernia.Impact statementWhat is already known on this subject? Laparoscopy has been used for the last 30 years. Constant improvement in the technique and equipment has allowed extensive, laparoscopic pelvic and abdominal surgery affording better outcomes than open surgery, an improved recovery, less pain, and fewer postoperative complications. However, mass removal remains a concern. Most laparoscopic specimens are larger than the initial trocar incision. Minimally invasive, adnexal mass surgery usually requires a trocar at least 10 mm wide to remove the mass. Alternatively, adnexal mass extraction from the abdominal cavity can proceed via a suprapubic, umbilical, or vaginal route.What do the results of this study add? During laparoscopic surgery, ovarian mass removal through an umbilical port using an endobag made from a surgical glove is useful due to the method requiring little funds, is easy to do, and results in a lower amount of postoperative pain than a removal via a lateral port using a standard endobag.What are the implications of these findings for clinical practice and/or further research? A transumbilical route using a bag made from a surgical glove is easy, economical, and causes less postoperative pain to the patient than removal via a lateral port employing a standard endobag.


[The surgical treatment of pancreatic pseudocyst].
Experience of surgical treatment of 370 patients with chronic pancreatitis, complicated by pancreatic pseudocyst occurrence, was analysed. Differentiational approach to operative intervention permitted to reduce the frequency of conduction of resections by 21.5%, replacing them by enhancing the quantity of drainage operations, what permitted to lower the occurrence of postoperative complications and to improve late follow-up result of treatment.


Lymphohematopoietic cancers in the United Farm Workers of America (UFW), 1988-2001.
Agricultural risk factors for lymphohematopoietic cancers (LHC) in Hispanic farm workers in California were examined in a nested case-control study embedded in a cohort of 139,000 ever members of a farm worker labor union in California. Crop and pesticide exposures were estimated by linking county/month and crop specific job history information from union records with California Department of Pesticide Regulation pesticide use reports during the 20-year period prior to cancer diagnosis. A total of 131 LHC diagnosed in California between 1988 and 2001 were included in the analysis. Analyses were conducted by gender and subtype of non-Hodgkins lymphoma (nodal, extra nodal) and by leukemia histology (lymphocytic, granulocytic). Odds ratios were calculated by stratification and by unconditional logistic regression. Risk for all LHC was elevated in workers cultivating vegetables (OR = 1.67, 95% CI = 1.12-2.48). Risk of leukemia was associated with exposure to the pesticides mancozeb (OR = 2.35, 95% CI = 1.12-4.95) and toxaphene (OR = 2.20, 95% CI = 1.04-4.65) while NHL risk was increased in association with 2,4-D (OR = 3.80, 95% CI=1.85-7.81). Risk of leukemia was particularly elevated among female workers and for granulocytic versus lymphocytic leukemia for several chemicals. No associations were noted for multiple myeloma. California farm workers employed where mancozeb and toxaphene were used had an increased risk of leukemia compared to farm workers employed elsewhere. Employment in farms using 2,4-D was associated with an increased risk of NHL.


Agreement Between StepWatch3 and ActiGraph wGT3X+ for Measuring Step-Based Metrics in People With Peripheral Artery Disease.
The authors investigated the agreement between StepWatch3™ (SW3) and ActiGraph™ wGT3X+ monitors for measuring step-based metrics in patients with peripheral artery disease and older adults. In 23 patients with peripheral artery disease and 38 older participants, the authors compared the metrics obtained during an outdoor (400-m track) walking session (step count) and a 7-day free-living period (step count and 60/30/5/1-min maximal or peak step accumulation) using the SW3 (ankle) and the wGT3X+ (hip) with the low-frequency extension filter enabled (wGT3X+/LFE) or not (wGT3X+/N). During outdoor walking session, agreement was high, particularly for wGT3X+/LFE: correlations ≥.98, median absolute percentage errors <1%, and significant equivalence using a ± 15% equivalence zone or narrower. In free living, no wGT3X+ method was equivalent to SW3 for step count. The wGT3X+/LFE was equivalent to SW3 regarding all step accumulation metrics using a ± 20% equivalence zone or narrower, with median absolute percentage errors <11%. The wGT3X+/LFE method is the best option for comparisons with SW3 in peripheral artery disease and older adults.


Coordinated regulation of transcription by CcpA and the Staphylococcus aureus two-component system HptRS.
The success of Staphylococcus aureus as a pathogen is due in part to its ability to adapt to changing environmental conditions using signal transduction pathways, such as metabolite- responsive regulators and two-component systems. S. aureus has a two-component system encoded by the gene pair sav0224 (hptS) and sav0223 (hptR) that regulate the hexose phosphate transport (uhpT) system in response to extracellular glucose-6-phosphate. Glycolytic intermediates such as glucose-6-phosphate are important carbon sources that also modulate the activity of the global metabolite-responsive transcriptional regulator CcpA. Because uhpT has a putative CcpA binding site in its promoter and it is regulated by HptR, it was hypothesized the regulons of CcpA and HptR might intersect. To determine if the regulatory domains of CcpA and HptRS overlap, ccpA was deleted in strains SA564 and SA564-ΔhptRS and growth, metabolic, proteomic, and transcriptional differences were assessed. As expected, CcpA represses hptS and hptR in a glucose dependent manner; however, upon CcpA derepression, the HptRS system functions as a transcriptional activator of metabolic genes within the CcpA regulon. Importantly, inactivation of ccpA and hptRS altered sensitivity to fosfomycin and ampicillin in the absence of exogenous glucose-6-phosphate, indicating that both CcpA and HptRS modulate antibiotic susceptibility.


Prosthetic and Surgical Management of Atypical Space When Teeth Are Missing.
When a patient presents with congenitally missing teeth, early diagnosis and comprehensive treatment planning are critical to effective restorative management. Interdental space allocation must be identified to accommodate proper clinical crown proportion(s) through a surgical-prosthetic solution. This article, which presents two case reports describing situations that clinicians may commonly face, demonstrates the management of atypical tooth spacing caused by congenitally missing teeth. The implementation of interdisciplinary therapy resulted in successful outcomes from both functional and esthetic perspectives.


Impact of diabetes mellitus on worsening of the left ventricular ejection fraction in exercise-gated 201Tl myocardial single photon emission computed tomography in patients with coronary artery disease.
It remains uncertain whether factors other than the severity of coronary artery disease (CAD) are associated with the worsening of the left ventricular ejection fraction (LVEF) by exercise. In the present study the impact of coronary risk factors on the worsening of LVEF by exercise was investigated in 391 patients with known or suspected CAD using exercise-gated (201)Tl scanning to calculate the LVEF. Significant worsening of the LVEF by exercise was defined as >4.7% (mean plus 1 SD of the value in 116 patients without CAD). Multivariate analysis revealed that diabetes mellitus (DM) was an independent risk factor for the worsening of LVEF by exercise in patients with multivessel (2- or 3-vessel) CAD with an odds ratio (95% confidence interval) of 2.2 (1.1-4.5, p=0.037). In 157 patients with 2- or 3-vessel CAD, 20 (23.5%) of 85 nondiabetic patients and 31 (43.1%, p=0.009 vs nondiabetic patients) of 72 diabetic patients showed significant worsening of LVEF by exercise. In patients with 2- or 3-vessel CAD, there was no significant difference in Gensini score or reversibility of perfusion defects between nondiabetic and diabetic patients. Thus, DM is a risk factor for worsening LVEF by exercise in addition to the severity of CAD.


Prevalence of respiratory symptoms in under 5s: 1993 to 2001.
To describe changes in the prevalence of respiratory symptoms in 1-4 year olds in two general practice populations observed on four occasions over an eight year period. In 1993, 1995, 1999, and 2001, questionnaires were posted to the parents of patients aged 15 years or younger and registered with either of two general practices. Only children aged 1-4 years at time of questionnaire completion were included in this study. For each survey, the prevalence of five key variables was determined. The response rates for all children in the four surveys were 72.8%, 70.6%, 65.0%, and 60.7% respectively. When respondents aged 1-4 years old were stratified into one-year age bands, there was a decrease in the prevalence of symptoms over the study period. This was statistically significant for wheeze and night cough in 2 year olds and for night cough in 4 year olds. Repeated antibiotic prescriptions decreased significantly for 2 and 3 year olds. There were no changes in the prevalence of hay fever or eczema and family history of asthma. The downward trend in symptom prevalence might represent a real decrease in symptoms or improvements in treatment. In the absence of changes in the prevalence of hay fever and family history of asthma, the downward trend in symptom prevalence may suggest changes in the prevalence of conditions other than asthma.


Isolation and characterization of microsatellite loci for the isopod crustacean Armadillidium vulgare and transferability in terrestrial isopods.
Armadillidium vulgare is a terrestrial isopod (Crustacea, Oniscidea) which harbors Wolbachia bacterial endosymbionts. A. vulgare is the major model for the study of Wolbachia-mediated feminization of genetic males in crustaceans. As a consequence of their impact on host sex determination mechanisms, Wolbachia endosymbionts are thought to significantly influence A. vulgare evolution on various grounds, including population genetic structure, diversity and reproduction strategies. To provide molecular tools for examining these questions, we isolated microsatellite loci through 454 pyrosequencing of a repeat-enriched A. vulgare genomic library. We selected 14 markers and developed three polymorphic microsatellite multiplex kits. We tested the kits on two A. vulgare natural populations and found high genetic variation, thereby making it possible to investigate the impact of Wolbachia endosymbionts on A. vulgare nuclear variation at unprecedented resolution. In addition, we tested the transferability of these kits by cross-species amplification in five other terrestrial isopod species harboring Wolbachia endosymbionts. The microsatellite loci showed good transferability in particular in Armadillidium nasatum and Chaetophiloscia elongata, for which these markers represent promising tools for future genetic studies.


The mouse fetal-placental arterial connection: A paradigm involving the primitive streak and visceral endoderm with implications for human development.
In Placentalia, the fetus depends upon an organized vascular connection with its mother for survival and development. Yet, this connection was, until recently, obscure. Here, we summarize how two unrelated tissues, the primitive streak, or body axis, and extraembryonic visceral endoderm collaborate to create and organize the fetal-placental arterial connection in the mouse gastrula. The primitive streak reaches into the extraembryonic space, where it marks the site of arterial union and creates a progenitor cell pool. Through contact with the streak, associated visceral endoderm undergoes an epithelial-to-mesenchymal transition, contributing extraembryonic mesoderm to the placental arterial vasculature, and to the allantois, or pre-umbilical tissue. In addition, visceral endoderm bifurcates into the allantois where, with the primitive streak, it organizes the nascent umbilical artery and promotes allantoic elongation to the chorion, the site of fetal-maternal exchange. Brachyury mediates streak extension and vascular patterning, while Hedgehog is involved in visceral endoderm's conversion to mesoderm. A unique CASPASE-3-positive cell separates streak- and non-streak-associated domains in visceral endoderm. Based on these new insights at the posterior embryonic-extraembryonic interface, we conclude by asking whether so-called primordial germ cells are truly antecedents to the germ line that segregate within the allantois, or whether they are placental progenitor cells. Incorporating these new working hypotheses into mutational analyses in which the placentae are affected will aid understanding a spectrum of disorders, including orphan diseases, which often include abnormalities of the umbilical cord, yolk sac, and hindgut, whose developmental relationship to each other has, until now, been poorly understood. This article is categorized under: Birth Defects > Associated with Preimplantation and Gastrulation Early Embryonic Development > Gastrulation and Neurulation.


Postmenopausal hyperandrogenism.
Postmenopausal hyperandrogenism is a state of relative or absolute androgen excess originating from the adrenal glands and/or ovaries clinically manifested by the presence of terminal hair in androgen-dependent areas of the body, and other manifestations of hyperandrogenism such as acne and alopecia or the development of virilization. In such circumstances, physicians must exclude the possibility of rare but serious androgen-producing tumors of the adrenal glands or ovaries. Worsening of undiagnosed hyperandrogenic disorders such as polycystic ovary syndrome, congenital adrenal hyperplasia, ovarian hyperthecosis, Cushing syndrome and iatrogenic hyperandrogenism should be considered for differential diagnosis. Elevated serum testosterone not only causes virilizing effects, but also will lead to hypercholesterolemia, insulin resistance, hypertension and cardiac disease. An ovarian androgen-secreting tumor, which is diagnosed in 1-3 of 1000 patients presenting with hirsutism, comprises less than 0.5% of all ovarian tumors. Adrenal tumors, including non-malignant adenomas and malignant carcinomas, are less common than ovarian tumors but cause postmenopausal virilization. Measurement of serum testosterone, sex hormone-binding globulin, dehydroepiandrosterone sulfate, androstenedione and inhibin B is necessary in postmenopausal women with the complaints and signs of hyperandrogenism. Some tests to discard Cushing syndrome should also be done. After an etiological source of androgen hypersecretion has been suspected, we recommend performing magnetic resonance imaging of the adrenal glands or ovaries. Medical management with gonadotropin-releasing hormone agonist/analogues or antagonists has been reported for women who are either unfit for surgery or in whom the source of elevated testosterone is unidentified.


The Bur1 cyclin-dependent kinase regulates telomere length in Saccharomyces cerevisiae.
Telomere length regulation is essential for cell viability in eukaryotes. While many pathways that affect telomere length are known, we do not yet have a complete understanding of the mechanism of length regulation. To identify new pathways that might regulate telomere length, we carried out a genetic screen in yeast and identified the cyclin-dependent kinase complex Bur1/2 as a regulator of telomere length. Mutations in either BUR1 cyclin-dependent kinase or the associated BUR2 cyclin resulted in short telomeres. This regulation did not function through the known role of BUR1 in regulating histone modification as bur1∆ set2∆ and bur2∆ set2∆ double mutants rescued cell growth but did not rescue the telomere shortening effects. We found that both bur1∆ and bur2∆ set2∆ were also defective in de novo telomere addition, and deletion of SET2 did also not rescue this elongation defect. The Bur1/2 cyclin-dependent kinase regulates transcription of many genes. We found that TLC1 RNA levels were reduced in bur2∆ set2∆ mutants; however, overexpression of TLC1 restored the transcript levels but did not restore de novo telomere elongation or telomere length. These data suggest that the Bur1/2 kinase plays a role in telomere elongation separate from its role in transcription of telomerase components. Dissecting the role of the Bur1/2 kinase pathway at telomeres will help complete our understanding of the complex network of telomere length regulation.


Contribution of endogenous glucagon-like peptide-1 to changes in glucose metabolism and islet function in people with type 2 diabetes four weeks after Roux-en-Y gastric bypass (RYGB).
Glucagon-Like Peptide-1 (GLP-1) is an insulin secretagogue which is elevated after Roux-en-Y Gastric Bypass (RYGB). However, its contribution to glucose metabolism after RYGB remains uncertain. We tested the hypothesis that GLP-1 lowers postprandial glucose concentrations and improves β-cell function after RYGB. To address these questions we used a labeled mixed meal to assess glucose metabolism and islet function in 12 obese subjects with type 2 diabetes studied before and four weeks after RYGB. During the post-RYGB study subjects were randomly assigned to receive an infusion of either saline or Exendin-9,39 a competitive antagonist of GLP-1 at its receptor. Exendin-9,39 was infused at 300 pmol/kg/min for 6 h. All subjects underwent RYGB for medically-complicated obesity. Exendin-9,39 resulted in increased integrated incremental postprandial glucose concentrations (181 ± 154 vs. 582 ± 129 mmol per 6 h, p = 0.02). In contrast, this was unchanged in the presence of saline (275 ± 88 vs. 315 ± 66 mmol per 6 h, p = 0.56) after RYGB. Exendin-9,39 also impaired β-cell responsivity to glucose but did not alter Disposition Index (DI). These data indicate that the elevated GLP-1 concentrations that occur early after RYGB improve postprandial glucose tolerance by enhancing postprandial insulin secretion.


High Precision DNA Modification Analysis of HCG9 in Major Psychosis.
New epigenetic technologies may uncover etiopathogenic mechanisms of major psychosis. In this study, we applied padlock probe-based ultra-deep bisulfite sequencing for fine mapping of modified cytosines of the HLA complex group 9 (nonprotein coding) gene in the postmortem brains of individuals affected with schizophrenia or bipolar disorder and unaffected controls. Significant differences between patients and controls were detected in both CpG and CpH modifications. In addition, we identified epigenetic age effects, DNA modification differences between sense and anti-sense strands, and demonstrated how DNA modification data can be used in clustering of patient populations. Our findings revealed new epigenetic complexities but also highlighted the potential of DNA modification approaches in the search of heterogeneous causes of major psychiatric disease.


Magnetophoresis 'meets' viscoelasticity: deterministic separation of magnetic particles in a modular microfluidic device.
The deflection of magnetic beads in a microfluidic channel through magnetophoresis can be improved if the particles are somehow focused along the same streamline in the device. We design and fabricate a microfluidic device made of two modules, each one performing a unit operation. A suspension of magnetic beads in a viscoelastic medium is fed to the first module, which is a straight rectangular-shaped channel. Here, the magnetic particles are focused by exploiting fluid viscoelasticity. Such a channel is one inlet of the second module, which is a H-shaped channel, where a buffer stream is injected in the second inlet. A permanent magnet is used to displace the magnetic beads from the original to the buffer stream. Experiments with a Newtonian suspending fluid, where no focusing occurs, are carried out for comparison. When viscoelastic focusing and magnetophoresis are combined, magnetic particles can be deterministically separated from the original streamflow to the buffer, thus leading to a high deflection efficiency (up to ~96%) in a wide range of flow rates. The effect of the focusing length on the deflection of particles is also investigated. Finally, the proposed modular device is tested to separate magnetic and non-magnetic beads.


Phosphosilicate fiber-based dual-wavelength random fiber laser with flexible power proportion and high spectral purity.
Phosphosilicate fiber has the inherent advantage of generating dual-wavelength output owing to the two Raman gain peaks at the frequency shifts of ∼13.2 THz (silica-related) and 39.9 THz (phosphorus-related), respectively. The frequency shift of 39.9 THz is often adopted to obtain long wavelength laser, while the control of Stokes light at 13.2 THz has attracted much attention currently. In this paper, a dual-wavelength random distributed feedback Raman fiber laser (RDFL) with over 100 nm wavelength interval and continuously tunable power proportion was presented based on phosphosilicate fiber for the first time. Through using the filtered amplified spontaneous emission (ASE) source as the pump source, the spectral purity of the Stokes light could be as high as 99.8%. By tuning two manual variable optical attenuators (VOAs), the power proportion of the silica-related Stokes light could range from ∼0% to 99.0%, and the maximum value is limited by the generation of second order Stokes light. Although the power handling capability of the VOA is merely 2 W, over 23 W total output power of the Stokes light was obtained thanks to the particular power distribution property of RDFL. This experiment demonstrates the potential to achieve a flexible high-power and high-spectral purity dual-wavelength RDFL output.


Transaxillary latissimus dorsi flap reconstruction in head and neck cancer. Limitations and refinements in 56 cases.
The transaxillary latissimus dorsi musculocutaneous flap is suitable whenever a large volume of tissue is required for head and neck reconstruction. Fifty-six transaxillary latissimus dorsi musculocutaneous flap reconstructions were performed in 55 patients. There were two cases of complete flap necrosis and eight cases of partial flap necrosis. The latissimus dorsi vascular pedicle is separate from the irradiated field. The pedicled latissimus dorsi flap provides coverage of the orbitocranium, including the supraorbital region and central portion of the upper face. In the event that the pedicled latissimus dorsi flap does not reach far enough cephalad, the nutrient vessels may be separated from the axillary artery and anastomosed to vessels in the neck. Combined defects of the esophagus, mandibulofacial region, and neck may be reconstructed with a single large latissimus dorsi flap. Hairless skin particularly suitable for oral cavity reconstruction is usually available. Aesthetic and functional deficits are minimal after latissimus dorsi reconstruction. Disadvantages of this technique include repositioning of the patient, increased blood loss, and longer operating time. Permanent brachial plexus injury may occur. The latissimus dorsi musculocutaneous flap should not be used when defects can be reconstructed by simpler methods.


Association with reward negatively modulates short latency phasic conditioned responses of dorsal raphe nucleus neurons in freely moving rats.
The dorsal raphe nucleus (DRN) is implicated in mood regulation, control of impulsive behavior, and in processing aversive and reward-related signals. DRN neurons show phasic responses to sensory stimuli, but whether association with reward modulates these responses is unknown. We recorded DRN neurons from rats in a contextual conditioned approach paradigm in which an auditory cue was either followed or not followed by reward, depending on a global context signal. Conditioned approach (licking) occurred after cues in the reward context, but was suppressed in the no-reward context. Many DRN neurons showed short-latency phasic activations in response to the cues. There was striking contextual bias, with more and stronger excitations in the no-reward context than in the reward context. Therefore, DRN activity scaled inversely with cue salience and with the probability of subsequent conditioned approach. Tonic changes were similarly discriminatory, with increases being dominant after cues in the no-reward context, when licking was suppressed, and tonic decreases in rate dominant after reward-predictive cues during expression of conditioned licking. Phasic and tonic DRN responses thus provide signals of consistent valence but over different timescales. The tonic changes in activity are consistent with previous data and hypotheses relating DRN activity to response suppression and impulse control. Phasic responses could contribute to this via online modulation of attention allocation through projections to sensory-processing regions.


[Ultrastructure of fat cells of obese patients with diabetes mellitus before and after treatment with a reducing diet].
Ultrastructural investigation of lipocytes of obese diabetics (Diabetes mellitus) before and after treatment with a reducing diet 21 diabetics not depending on insulin with obesity of the second or third degree were given for 3 months a reducing diet containing 5110 kJ/day (86 g protein, 41 g fat, 119 g carbohydrates). After treatment the test persons had a loss of weight of 7.60 kg on average. Before and after treatment subcutaneous adipose tissue was taken by puncture biopsy and investigated by electron microscopy. After treatment extended intercellular spaces are evident in which collagenic and elastic fibres appear quantitatively enlarged. A certain plication and inequality of the cell membranes, a penetration of the fat droplet by cytoplasmatic outgrowths as well as a diminution of the lipocytes and the big fat droplet were found. The investigations show that even the relatively short treatment of 3 months exercises an influence on the structure of the lipocytes. That shows new possibilities of therapy and prophylaxis of obesity.


Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells.
Oct-3/4 is a key transcriptional factor whose expression level governs the fate of primitive inner cell mass and embryonic stem (ES) cells. Previously, an upstream 3.3-kb distal enhancer (DE) fragment was identified to be responsible for the specific expression of mouse Oct-3/4 in the inner cell mass and ES cells. However, little is known about the cis-elements and trans-factors required for DE activity. In this study, we identified a novel cis-element, called Site 2B here, located approximately 30 bp downstream from Site 2A, which was previously revealed in DE by an in vivo chemical modification experiment. Using the luciferase reporter assay, we demonstrated that both Site 2A and Site 2B are necessary and sufficient for activating DE in the contexts of both the native Oct-3/4 promoter and the heterologous thymidine kinase minimal promoter. In an electrophoretic mobility shift assay we showed that Site 2B specifically binds to Oct-3/4 and Sox2 when ES-derived cell extracts were used, whereas Site 2A binds to a factor(s) present in both ES and NIH 3T3 cells. Furthermore, we showed that the physiological level of Oct-3/4 in ES cells is required for Site 2B-mediated DE activity using the inducible knock-out system of Oct-3/4 in ES cells. These results indicate that Oct-3/4 is a member of the gene family regulated by Oct-3/4 and Sox2, as reported before for the FGF-4, UTF1, Sox2, and Fbx15 genes. Thus, Oct-3/4 and Sox2 comprise a regulatory complex that controls the expression of genes important for the maintenance of the primitive state, including themselves. This autoregulatory circuit of the Sox2.Oct-3/4 complex may contribute to maintaining robustly the precise expression level of Oct-3/4 in primitive cells.


Effectiveness of Respiratory Muscle Training on Respiratory Muscle Strength, Pulmonary Function, and Respiratory Complications in Stroke Survivors: A Systematic Review of Randomized Controlled Trials.
Stroke is the most common leading cause of mortality and related morbidities worldwide. After stroke, the motor function of extremities and spinal muscles is significantly impairment, but not only this, it also has attributable factors leading to respiratory dysfunction. Nevertheless, to the extent of the authors' knowledge, there is a dearth of conclusive studies which examined the effectiveness of RMT on muscle strength, pulmonary function, and respiratory complications of individuals after stroke. The purpose of this systematic review was to evaluate the effectiveness of respiratory muscle training on respiratory muscle strength, pulmonary function, and respiratory complications in patients after stroke. An electronic database search of HINARI, PEDro, PubMed, Cochrane Library and Google scholar was used to identify randomized controlled trials that evaluated the effectiveness of respiratory muscle training in patients with stroke. Articles published from 2010 to 2019 were included. The quality of the articles was assessed using PEDro scale. Articles with abstract only, PEDro scores less than 5, published in non-English language, not freely available articles, and quasi experimental studies were excluded from this study. The literature search yielded a total of 7 articles (6 randomized controlled trials with 1 pilot randomized controlled trial) which met inclusion criteria despite their heterogeneity. The methodological quality of all studies ranged from 6 to 8 in Pedro score. Most of the articles reported a significant increase in respiratory muscle strength, respiratory muscle function, and reduced risk of complications with a p value <0.05. Respiratory muscle training could potentially improve muscle strength and pulmonary functions of subjects after stroke. Thus, it may reduce stroke-related respiratory complications in subjects after stroke. However, further study is warranted with high quality RCTs and pooled synthesis of results.


Previous Emergency Medical Services Use by Victims of Child Homicide.
The medical diagnoses and frequency of emergency department visits made by children who are later given a diagnosis of maltreatment do not differ much from those of nonabused children. However, the type of medical complaints and frequency of emergency medical services (EMS) use by child homicide victims before their death are not known. We compared EMS use between child homicide victims and children who died from natural causes before their death. This was a retrospective case-control study of children 0 to 5 years old who died in Houston, Texas, from 2005 to 2010. Cases were child homicide victims. Controls were children who died from natural causes. We reviewed death data and EMS and child protective services (CPS) encounter information before the victim's death. The association between death type (natural vs homicide) and EMS use was assessed using Poisson regression with EMS count adjusted for exposure time. There were 89 child homicides and 183 natural deaths. Age at death was significantly higher for homicides than natural deaths (1.1 vs 0.2 y, P < 0.001). Homicide victims used EMS services (39% vs 14%, P < 0.001) and had previous CPS investigations (55% vs 7%, P < 0.001) significantly more often than children who died from natural causes. Poisson regression, after adjustment for age, revealed that the homicide group had more EMS calls than the natural death group (β = 0.55; 95% confidence interval, 0.04-1.07; P = 0.03). However, the EMS use frequency and working assessments were not helpful in identifying maltreatment victims. Child homicide victims use EMS more often and have a higher number of CPS investigations before their death than children who die from natural causes. However, the frequency and nature of EMS medical complaints are not helpful in identifying maltreatment.


Natural history of Sin Nombre virus in western Colorado.
A mark-recapture longitudinal study of immunoglobulin G (IgG) antibody to Sin Nombre virus (SNV) in rodent populations in western Colorado (1994-results summarized to October 1997) indicates the presence of SNV or a closely related hantavirus at two sites. Most rodents (principally deer mice, Peromyscus maniculatus, and pinyon mice, P. truei) did not persist on the trapping webs much beyond 1 month after first capture. Some persisted more than 1 year, which suggests that even a few infected deer mice could serve as transseasonal reservoirs and mechanisms for over-winter virus maintenance. A positive association between wounds and SNV antibody in adult animals at both sites suggests that when infected rodents in certain populations fight with uninfected rodents, virus amplification occurs. At both sites, male rodents comprised a larger percentage of seropositive mice than recaptured mice, which suggests that male mice contribute more to the SNV epizootic cycle than female mice. In deer mice, IgG antibody prevalence fluctuations were positively associated with population fluctuations. The rates of seroconversion, which in deer mice at both sites occurred mostly during late summer and midwinter, were higher than the seroprevalence, which suggests that the longer deer mice live, the greater the probability they will become infected with SNV.


CRIB (NGF delivery) Cyto Therapeutics Inc.
Cellular Replacement by Immunoisolatory Biocapsule (CRIB) is a technology under development by Cyto-Therapeutics for the delivery of therapeutic substances to the central nervous system (CNS). Preclinical studies with implants are underway for the potential treatment of Alzheimer's disease, Huntington's chorea and other neurodegenerative diseases [180367]. Theses implants contain cells that have been genetically engineered to secrete human nerve growth factor. Development of the CRIB implant is in collaboration with Genentech. CRIB consists of product-secreting cells encapsulated in a semipermeable polymer membrane. The device is surgically implanted into the target area of the brain, thus bypassing the blood-brain barrier, whilst isolating the foreign cells from the patient's immune system [180367]. CRIB implants secreting nerve growth factor (NGF) were effective in preventing nerve degeneration in a monkey model of Alzheimer's disease. The devices contained baby hamster kidney fibroblasts (genetically modified to secrete human NGF) and were implanted into brain ventricles of elderly rhesus monkeys whose cholinergic neurons had been cut. The monkeys treated with non-NGF secreting implants showed a decrease in the number of cholinergic neurons, while those treated with NGF-secreting implants showed a smaller loss of neurons and sprouting of cholinergic nerve fibres [200261]. CytoTherapeutics is working towards clinical trials with these implants in patients with Alzheimer's disease [168581]. Trials of NGF-secreting implants have also been carried out in animal models of Huntington's disease. The company demonstrated that in rodents whose brains had been lesioned with quinolinic acid to approximate the neuronal death which occurs in the disease, NGF-releasing implants protected many striatal neurons that would normally have died. In addition, it was shown that behaviors normally lost as a result of striatal neuron damage could be significantly attenuated. Further investigations are scheduled for the potential use of CRIB/NGF technology in Huntington's disease [180367]. The CRIB technology was originally developed at Brown University (Rhode Island, USA) which owns several patents that cover the technology. CytoTherapeutics has entered into collaborative agreements with the university for the use of the technology [199903] as well as with NeuroSpheres, Canada, for the use of their neural stem cells [199898].


Structurally stabilized organosilane-templated thermostable mesoporous titania.
Structurally thermostable mesoporous anatase TiO2 (m-TiO2) nanoparticles, uniquely decorated with atomically dispersed SiO2, is reported for the first time. The inorganic Si portion of the novel organosilane template, used as a mesopores-directing agent, is found to be incorporated in the pore walls of the titania aggregates, mainly as isolated sites. This is evident by transmission electron microscopy and high-angle annular dark field scanning transmission electron microscopy, combined with electron dispersive X-ray spectroscopy. This type of unique structure provides exceptional stability to this new material against thermal collapse of the mesoporous structure, which is reflected in its high surface area (the highest known for anatase titania), even after high-temperature (550 °C) calcination. Control of crystallite size, pore diameter, and surface area is achieved by varying the molar ratios of the titanium precursor and the template during synthesis. These mesoporous materials retain their porosity and high surface area after template removal and further NaOH/HCl treatment to remove silica. We investigate their performance for dye-sensitized solar cells (DSSCs) with bilayer TiO2 electrodes, which are prepared by applying a coating of m-TiO2 onto a commercial titania (P25) film. The high surface area of the upper mesoporous layer in the P25-m-TiO2 DSSC significantly increases the dye loading ability of the photoanode. The photocurrent and fill factor for the DSSC with the bilayer TiO2 electrode are greatly improved. The large increase in photocurrent current (ca. 56%) in the P25-m-TiO2 DSSC is believed to play a significant role in achieving a remarkable increase in the photovoltaic efficiency (60%) of the device, compared to DSSCs with a monolayer of P25 as the electrode.


Selection of a Biosafety Level 1 (BSL-1) surrogate to evaluate surface disinfection efficacy in Ebola outbreaks: Comparison of four bacteriophages.
The 2014 West African Ebola virus disease outbreak was the largest to date, and conflicting, chlorine-based surface disinfection protocols to interrupt disease transmission were recommended. We identified only one study documenting surface disinfection efficacy against the Ebola virus, showing a >6.6 log reduction after 5-minute exposure to 0.5% sodium hypochlorite (NaOCl) based on small-scale tests (Cook et al. (2015)). In preparation for future extensive, large-scale disinfection efficacy experiments, we replicated the Cook et al. experiment using four potential BSL-1 surrogates selected based on similarities to the Ebola virus: bacteriophages MS2, M13, Phi6, and PR772. Each bacteriophage was exposed to 0.1% and 0.5% NaOCl for 1, 5, and 10 minutes on stainless steel. MS2 and M13 were only reduced by 3.4 log and 3.5 log after a 10-minute exposure to 0.5% NaOCl, and would be overly conservative surrogates. Conversely, PR772 was too easily inactivated for surrogate use, as it was reduced by >4.8 log after only 1-minute exposure to 0.5% NaOCl. Phi6 was slightly more resistant than the Ebola virus, with 4.1 log reduction after a 5-minute exposure and not detected after a 10-minute exposure to 0.5% NaOCl. We therefore recommend Phi6 as a surrogate for evaluating the efficacy of chlorine-based surface disinfectants against the Ebola virus.


A multiple imputation score test for model modification in structural equation models.
Structural equation modeling (SEM) applications routinely employ a trilogy of significance tests that includes the likelihood ratio test, Wald test, and score test or modification index. Researchers use these tests to assess global model fit, evaluate whether individual estimates differ from zero, and identify potential sources of local misfit, respectively. This full cadre of significance testing options is not yet available for multiply imputed data sets, as methodologists have yet to develop a general score test for this context. Thus, the goal of this article is to outline a new score test for multiply imputed data. Consistent with its complete-data counterpart, this imputation-based score test provides an estimate of the familiar expected parameter change statistic. The new procedure is available in the R package semTools and naturally suited for identifying local misfit in SEM applications (i.e., a model modification index). The article uses a simulation study to assess the performance (Type I error rate, power) of the proposed score test relative to the score test produced by full information maximum likelihood (FIML) estimation. Due to the two-stage nature of multiple imputation, the score test exhibited slightly lower power than the corresponding FIML statistic in some situations but was generally well calibrated. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Highly structured sequence homology between an insertion element and the gene in which it resides.
The recessive allele for soybean seed lectin results from the insertion of a DNA segment (designated Tgm1) into the coding region of the gene. The termini of Tgm1 display structural features characteristic of a transposable element. The complete sequence of Tgm1 contains 3550 base pairs (bp) and can be divided into three regions (left arm, mid-section, and right arm). No large open reading frames were found, but an extensive, highly structured border with homology to the lectin gene was revealed. The left border (726 bp) comprising most of the left arm and extreme right border (144 bp) of the right arm consist of various forms of a basic 54-bp repeating unit. This 54-bp unit is comprised of a stem-loop structure and interhairpin sequence that occurs 13 times in the left arm and 2 times in the right arm of Tgm1. Progressively degenerate forms of this repeating unit appear toward the termini of Tgm1, but the dyad symmetry remains highly conserved. Seven nucleotides (A-C-A-T-C-G-G and its complement) maintained within the stem also appear as a subset of inverted repeats found at nearly equal distances from the target site in the lectin gene. Together with the inverted repeat termini and a duplication in the left arm, this 7-bp sequence occurs a total of 33 times in Tgm1. We infer that the dyad symmetries containing this sequence are involved in target gene selection. The repeating unit format of Tgm1 describes a distinct class of eukaryotic elements that includes representatives known to be mobile in snapdragon and maize.


Prognostic factors in the treatment of mild hypertension. The Management Committee of the Australian National Blood Pressure Study.
In the Australian therapeutic trial in patients with mild diastolic hypertension, univariate analysis of possible prognostic characteristics (covariates) at the time of entry into the study showed a higher incidence of trial end points, mainly events due to ischemic heart disease and cerebrovascular disease, in older subjects, men, cigarette smokers, and those with higher systolic blood pressures and lower body mass indexes than in their complementary groups. There was a lower incidence of these events in actively treated subjects than in control subjects at both high and low levels of incidence of each covariate considered, but treatment benefit was most marked in those with lower serum cholesterol levels and lower systolic blood pressures. Multivariate regression analyses of data from men and women separately, performed with the Cox proportional-hazards model, confirmed that the incidence of trial end points increased with age and with systolic blood pressure, and showed higher rates in cigarette smokers, which increased markedly with decreasing body weight. With respect to the effects of treatment, the multivariate analysis showed an increasing benefit with decreasing body weight in smokers. The apparent relationship of treatment benefit to systolic blood pressure in the univariate analysis did not reach statistical significance in the multivariate analysis. The greater benefit from treatment related to lower levels of serum cholesterol found in the univariate analysis was also found in several preliminary multivariate analyses, but did not reach statistical significance in the final model. However, this relationship should be tested in other therapeutic trials in progress or recently completed.(ABSTRACT TRUNCATED AT 250 WORDS)


Epidermoid cyst in an intrapancreatic accessory spleen: a case report.
Ectopic splenic tissue in the abdominal cavity is a common entity, with a reported incidence of 10% in the general population. However, an intrapancreatic accessory spleen is a rare disease, and moreover cyst formation in it is exceedingly rare. A 58-year-old woman with a 25-mm multilocular cyst in the tail of the pancreas detected incidentally by ultrasonography was admitted for further evaluation. Because malignancy could not be ruled out, a spleen-preserving distal pancreatectomy was performed. The cut surface of the surgical specimen showed a multilocular cyst surrounded by brown solid tissue resembling normal spleen. Pathological examination revealed it was stratified squamous epithelium and was surrounded by splenic tissue. The final pathological diagnosis was epidermoid cyst in an accessory spleen in the pancreas. This cyst has no characteristic features on diagnostic imaging. Consequently, it is not possible to make a definite preoperative diagnosis in most cases. Epidermoid cyst in intrapancreatic splenic tissue is another lesion to be considered in the differential diagnosis of pancreatic tail tumors.


Localization model description of diffusion and structural relaxation in superionic crystalline UO2.
We test the Localization Model (LM) prediction of a parameter-free relationship between the α-structural relaxation time τα and the oxygen ion diffusion coefficient DO with the Debye-Waller factor ⟨u2⟩ for crystalline UO2 under superionic conditions where large anharmonic interactions lead to non-Arrhenius relaxation and high ion mobility. As in a previous study of structural relaxation in Cu-Zr metallic glass materials having a range of compositions, we find that the LM relationship between the picosecond atomic dynamics ("fast" beta relaxation) and the long-time structural relaxation time and oxygen ion diffusion coefficient holds to an excellent approximation without any free parameters over the full range of temperatures and pressures investigated in our simulations.


Prevalence of Mycoplasma pneumoniae Infection in Malagasy Children.
Childhood community-acquired pneumonia is a leading cause of childhood morbidity in low-income countries. The etiologic agents are usually Staphylococcus aureus, Streptococcus pneumoniae and Mycoplasma pneumoniae. M. pneumoniae was recognized as a cofactor in asthmatic disease. High asthma prevalence was reported in Madagascar. Our aim was to clarify the prevalence of M. pneumoniae infection in this country and its relationship with asthma. A prospective study was conducted in 351 children (from 2 to 16 years of age) from January 2012 to December 2014. According to the clinical symptoms, children were enrolled in 3 groups: "control group" (CG, n = 106), "asthma group" (n = 129) and "pneumonia group" (n = 116). The IgG and IgM M. pneumoniae status was evaluated by an enzyme-linked immunosorbent assay. Clinical signs of infection, socioeconomic data and antimicrobial treatment were recorded. The overall prevalence of M. pneumoniae infection was 18.2%. The multivariate analysis demonstrated that M. pneumoniae infection was significantly more frequent in the CG [pneumonia group vs. CG: odds ratio = 0.45 (0.21-0.91), P = 0.037 and asthma group vs. CG: odds ratio = 0.39 (0.18-0.87), P = 0.021]. The C-reactive protein value was significantly higher in children with M. pneumonia-positive serology (85 vs. 61 mg/L, P = 0.03). Of note, 99 (41%) children received antibiotics before attending. We report a prevalence of 18.2% for M. pneumoniae infection in children in Madagascar. The prevalence of M. pneumoniae infection was higher in the control patients than in asthmatic ones.


In vitro phenotypes to elvitegravir and dolutegravir in primary macrophages and lymphocytes of clonal recombinant viral variants selected in patients failing raltegravir.
The cross-resistance profiles of elvitegravir and dolutegravir on raltegravir-resistant variants is still controversial or not available in macrophages and lack extensive evaluations on wide panels of clonal variants. Thus, a complete evaluation in parallel with all currently available integrase inhibitors (INIs) was performed. The integrase coding region was RT-PCR-amplified from patient-derived plasma samples and cloned into an HIV-1 molecular clone lacking the integrase region. Twenty recombinant viruses bearing mutations to all primary pathways of resistance to raltegravir were phenotypically evaluated with each integrase inhibitor in freshly purified CD4+ T cells or monocyte-derived macrophages. Y143R single mutants conferred a higher level of raltegravir resistance in macrophages [fold change (FC) 47.7-60.24] compared with CD4+ T cells (FC 9.55-11.56). All other combinations had similar effects on viral susceptibility to raltegravir in both cell types. Elvitegravir displayed a similar behaviour both in lymphocytes and macrophages with all the tested patterns. When compared with raltegravir, none to modest increases in resistance were observed for the Y143R/C pathways. Dolutegravir maintained its activity and cross-resistance profile in macrophages. Only Q148H/R variants had a reduced level of susceptibility (FC 5.48-18.64). No variations were observed for the Y143R/C (+/-T97A) or N155H variants. All INIs showed comparable antiretroviral activity in both cell types even if single mutations were associated with a different level of susceptibility in vitro to raltegravir and elvitegravir in macrophages. In particular, dolutegravir was capable of inhibiting with similar potency infection of raltegravir-resistant variants with Y143 or N155 pathways in both HIV-1 major cell reservoirs.


Preparation of Nonprecious Metal Electrocatalysts for the Reduction of Oxygen Using a Low-Temperature Sacrificial Metal.
Non-platinum group metal (non-PGM) electrocatalysts for the oxygen reduction reaction (ORR) are generally composed of iron, nitrogen, and carbon synthesized through high-temperature pyrolysis. Among the various types of precursors, metal-organic frameworks (MOFs), zeolitic imidazolate framework (ZIF)-8 in particular, have often been used in the synthesis. The pyrolysis of ZIF-8 precursor relies on the use of Zn as a sacrificial metal (SM), and the optimal processing temperatures often exceed 1000 °C to generate active non-PGM catalysts. The high pyrolysis temperature tends to result in heterogeneous active moieties ranging from Fe single atoms to nanoparticles. In this study, we present the synthesis of non-PGM catalysts using Cd as the sacrificial metal instead of Zn. By using Cd, we were able to generate active non-PGM electrocatalysts from the MOF precursors at a low pyrolysis temperature of 750 °C, which helps preserve the single atomic iron active sites.


Cytokine Effect of TLR3, TLR4, and TLR7 Agonists Alone or Associated with Leishmania infantum Antigen on Blood from Dogs.
Activation of toll-like receptors (TLRs) has been shown to play an important role in leishmaniosis by enhancing the parasite specific immune responses to control infection. However, the role of TLR agonists has not been studied in detail in dogs. The aim of this study was to determine the effect of TLR3, TLR4, and TLR7 agonists (TLR3a, TLR4a, and TLR7a) alone or in combination with Leishmania infantum antigen (LSA) on TNF-α and IL-6 production in blood from dogs living in endemic areas of canine leishmaniosis (CanL). Twenty-four healthy dogs from Catalonia (n=14) and Ibizan hound dogs from the island of Mallorca (n=10) were enrolled. Whole blood with TLR3a, TLR4a, and TLR7a alone or combined with LSA were cultured separately, and IFN-γ, TNF-α, and IL-6 were measured by ELISA. A significant increase of TNF-α was found for all conditions studied compared to medium alone. Stimulation with TLR4a (p=0.0001) and TLR7a (p=0.005) presented a significantly marked increase in TNF-α and IL-6 production compared to TLR3a. Importantly, significantly higher TNF-α production was found in LSA+TLR4a (p=0.0001) stimulated blood and LSA+TLR7a (p=0.005) compared to LSA alone. All dogs showed higher TNF-α production after LSA+TLR7a compared to TLR7a (p=0.047) and LSA+TLR3a compared to TLR3a (p=0.052). These data indicate a marked inflammatory cytokine effect of TLR4a and TLR7a on blood from healthy dogs living in endemic areas of CanL. Additionally, LSA+TLR7a promoted a synergistic proinflammatory effect with TNF-α in all dogs. Those findings suggest an active role of TLRs in proinflammatory responses, which might be strongly involved in the process of disease resolution.


Spiral CT in gastric carcinoma: comparison with barium study, fiberoptic gastroscopy and histopathology.
To evaluate spiral computed tomography (CT) including virtual gastroscopy for diagnosis of gastric carcinoma in comparison with upper gastrointestinal series (UGI), fiberoptic gastroscopy (FG) and histopathology. Sixty patients with histologically proven gastric carcinoma (54 advanced and 6 early) were included in this study. The results of spiral CT were compared with those of UGI and FG. Two observers blindly evaluated images of spiral CT and UGI and video recording of FG with consensus in terms of diagnostic confidence with a five-point scale. Sensitivities of lesion detection, Borrmann's classification of spiral CT, UGI and FG, as well as the accuracy of TNM staging of spiral CT were determined by comparing them to surgical and histological findings. The lesion detection rate was 98 % (59/60), 95 % (57/60) and 98 % (59/60) for spiral CT, UGI and FG, respectively. There were no statistical differences in the detection sensitivity among the three techniques (P>0.05). For the sensitivity in Borrmann's classification, spiral CT was higher than that of UGI (P=0.025) and similar to that of FG (P>0.05). The accuracy of spiral CT in staging the gastric carcinoma was 76.7 %. Six cases of early gastric carcinoma were all detected by spiral CT as well as FG. Spiral CT is equivalent to UGI and FG in the detection of gastric carcinoma, and superior to UGI but similar to FG in the Borrmann's classification of advanced gastric carcinoma. Spiral CT is more valuable than FG in the staging of gastric carcinoma.


Nitric oxide inhibits stretch-induced MAPK activation in mesangial cells through RhoA inactivation.
Glomerular capillary hypertension is an important determinant of glomerulosclerosis in rats with subtotal renal ablation. Dietary supplementation with L-arginine increases renal nitric oxide (NO) production and limits glomerular injury in this model, and early benefits are seen without altered glomerular capillary pressure. In an in vitro model of hemodynamically mediated signaling, the authors have reported that subjecting MC to cyclic stretch/relaxation activates the mitogen-activated protein kinase p42/44 (Erk) cascade and that NO and cyclic GMP abrogate stretch-induced Erk activation by inducing actin cytoskeletal disassembly. The actin cytoskeleton is regulated by the Rho family of GTPases, including RhoA; therefore, the authors examined the role of RhoA in stretch-induced Erk activation and as an NO target. In primary rat MC subjected to cyclic mechanical strain, RhoA activity was maximally increased (2.4-fold) after 1 min of stretch, and Erk activation temporally followed. The Rho-kinase inhibitor Y-27632 attenuated Erk activation in a dose-dependent manner and prevented stretch-induced actin stress fiber formation. The NO donors S-nitroso-N-acetylpenicillamine and cGMP both inhibited stretch-induced RhoA and Erk activation and stress fiber formation. Infection of MC with the RhoA mutant RhoA-Ala188, which is resistant to NO-dependent phosphorylation, abrogated the effects of NO and cGMP on stretch-induced Erk activation and stress fiber formation. The authors conclude that the early activation of RhoA is essential for stretch-induced actin stress fiber formation and Erk activation in MC, events which are prevented by NO and cGMP through their action on RhoA. Inhibition of RhoA may thus be a new approach to the prevention of hemodynamically mediated glomerular injury.


Tarsodermal suture fixation preceding redundant skin excision: a modified non-incisional upper blepharoplasty method for elderly patients.
Non-incisional blepharoplasty is a simple, less invasive method for creating a more natural-appearing double eyelid than classical incisional blepharoplasty. However, in aging patients, non-incisional blepharoplasty is not effective due to more severe blepharochalasis. Traditionally, incisional blepharoplasty is a common surgical method used for older patients, but blepharoplasty in elderly patients typically results in prolonged recovery times, and final blepharoplasty lines may be located in unintended or asymmetrical positions. Here, we introduce a new modified combination technique for geriatric blepharoplasty. A total of ten patients were treated from July 2010 through July 2012 using the combination method. First, we performed non-incisional blepharoplasty using tarsodermal fixation. Then, incisional blepharoplasty with additional elliptical excision of the upper eyelid skin was performed. We removed pretarsal tissue, fat, the orbicularis oculi muscle, and orbital fat. Telephone surveys were administered to all patients for follow-up. The questionnaire was composed of eight questions that addressed recurrence and satisfaction with aesthetics and the procedure. A total of nine patients (90%) responded to the telephone survey. All cases of moderate to severe blepharochalasia were corrected and there were no major complications. Patients who underwent blepharoplasty had higher satisfaction scores. All patients were satisfied with the postoperative shapes of their eyelids. THE ADVANTAGES OF THE PROPOSED TECHNIQUE INCLUDE: ease of obtaining a natural-looking fold with symmetry at the desired point; reproducible methods that require short operation times; fast postoperative recovery that results in a natural-appearing double-eyelid line; and high patient satisfaction.


The structure and function of claudins, cell adhesion molecules at tight junctions.
Tight junctions (TJs) play a pivotal role in compartmentalization in multicellular organisms by sealing the paracellular pathway in epithelial and endothelial cell sheets. Recently, novel integral membrane proteins, claudins, have been identified as major cell adhesion molecules working at TJs. Claudins comprise a multigene family, and each member of approximately 23 kDa bears four transmembrane domains. To date, 15 members of this gene family have been identified. When expression vectors of each species of claudins were transfected into fibroblasts lacking endogenous claudins or TJs, well-developed TJs were observed between adjacent transfectants. Furthermore, claudins were shown to be directly involved in the barrier function of TJs by experiments using Clostridium perfringens enterotoxin. Now that claudins have been identified, the structure and functions of TJs should be determined in detail in molecular terms.


Mevalonic acid products as mediators of cell proliferation in simian virus 40-transformed 3T3 cells.
Effects of treatment with serum-free medium and 25-hydroxycholesterol (25-OH) on the cell cycle of simian virus 40-transformed 3T3 fibroblasts, designated SV-3T3 cells, were studied and compared with simultaneous effects on the activity of 3-hydroxy-3-methylglutaryl (HMG) CoA reductase and incorporation of [3H]mevalonic acid into cholesterol, Coenzyme Q, and dolichol. The data confirm our previous finding (O. Larsson and A. Zetterberg, Cancer Res., 46: 1233-1239, 1986) that 25-OH inhibits the cell cycle traverse of SV-3T3 cells specifically in early G1. In contrast, treatment with serum-free medium had no effect on cell cycle progression. The effect of 25-OH on the cell cycle traverse was correlated to a substantial decrease in the activity of HMG CoA reductase, whereas there was no change in the rate of [3H]mevalonic acid incorporated into cholesterol, Coenzyme Q, and dolichol. When the cells were exposed to serum-free medium, there was no depression of activity of HMG CoA reductase, and the rate of [3H]mevalonic acid incorporated into dolichol and cholesterol was not affected in any appreciable degree. In contrast the rate of Coenzyme Q synthesis was substantially decreased as a result of serum depletion. A similar decrease in Coenzyme Q synthesis was also achieved by treating the cells with cholesterol-poor serum. This indicates that the rate of Coenzyme Q synthesis is dependent on the concentration of cholesterol in the culture medium. In order to analyze whether some of the products in the mevalonic acid biosynthetic pathway may be of importance in the control of G1 traverse and cell proliferation of SV-3T3 cells, cholesterol, Coenzyme Q, and dolichol were added as supplements to cells treated with 25-OH. It was shown that dolichol was capable of overcoming the 25-OH-induced inhibition of G1 traverse efficiently, whereas cholesterol and Coenzyme Q were considerably less effective. Considered together with the fact that the activity of HMG CoA reductase and incorporation of mevalonic acid into dolichol were unaffected following serum-free treatment, the results suggest that maintenance of a certain level of de novo synthesis of dolichol may contribute to the capability of SV-3T3 cells to proliferate in serum-free medium.


Cell-Membrane Coated Self-Immolative Poly(thiourethane) for Cysteine/Homocysteine-Triggered Intracellular H2S Delivery.
Hydrogen sulfide (H2S) is an important gaseous signaling molecule with unique pleiotropic pharmacological effects, but may be limited for clinical translation due to the lack of a reliable delivery form that delivers exogenous H2S to cells at action site with precisely controlled dosage. Herein, we report the design of a poly(thiourethane) (PTU) self-immolative polymer terminally caged with an acrylate moiety to trigger release of H2S in response to cysteine (Cys) and homocysteine (Hcy), the most used and independent indicators of neurodegenerative diseases. The synthesized PTU polymer was then coated with the red-blood-cell (RBC) membrane in the presence of solubilizing agent to self-assemble into nanoparticles with enhanced stability and cytocompatibility. The Hcy/Cys mediated addition/cyclization chemistry actuated the biomimetic polymeric nanoparticles to disintegrate into carbonyl sulfide (COS), and finally convert into H2S via the ubiquitous carbonic anhydrase (CA). H2S released in a controlled manner exhibited a strong antioxidant ability to resist Alzheimer's disease (AD)-related oxidative stress factors in BV-2 cells, a neurodegenerative disease model in vitro. Thus, this work may provide an effective strategy to construct H2S donors that can degrade in response to a specific pathological microenvironment for the treatment of neurodegenerative diseases.


On closed extensions of functions.
G. T. Whyburn has shown that every mapping (by a mapping we will always mean a continuous function) of a topological space into a Hausdorff space is the restriction of a compact mapping with the same range (Trans. Am. Math. Soc., 74, 344-350 (1953)). H. Bauer has proved that every mapping between locally compact Hausdorff spaces can be extended in a topologically unique manner to a compact mapping on a locally compact Hausdorff space with the same range (Math. Ann., 138, 398-427 (1959)). When the range and domain of a mapping are locally compact Hausdorff spaces, Whyburn's and Bauer's extensions are topologically equivalent. It is well known that not every compact mapping is a closed mapping; however, Whyburn has shown that every closed function with compact point inverses is a compact function (these PROCEEDINGS, 54, 688-692 (1965)). The principal aim of this paper is to prove that every function (respectively, mapping) from one topological space to another can be extended in a topologically unique manner to a closed and compact function (respectively, mapping) with the same range.


Children's Pain Perception and Behavioral Feedback during Local Anesthetic Injection with Four Injection Site Preparation Methods.
Introduction:An effective pain control significantly contributes to an optimal dental treatment in pediatric dentistry. This study was conducted to compare children's pain perception and behavioral feedback during local anesthetic injection accompanied with counter-irritation, refrigerant, ice precooling or topical benzocaine. Methods:This study was conducted on 99 children who needed dental treatment in bilateral maxillary primary molars by local anesthesia. Subjects were randomly assigned to three groups, in which the injection site was prepared using counter-irritation in group I, ice precooling in group II, and refrigerant spray precooling in group III. In all three groups, 20% benzocaine gel was used in the injection site of opposite quadrant as a control. The perceived pain and behavioral feedback of children during injection were evaluated using the visual analogue scale (VAS) and sound, eye, and motor (SEM) indexes. Sign and Kruskal-Wallis tests were used to analyze data at a significance level of P < 0.05. Results:The perceived pain was significantly lower when using benzocaine compared to the other three methods (P < 0.05). Behavioral feedback of children when using benzocaine was not significantly different from the counter-irritation method (P=1.00). However, behavioral feedback with counter-irritation was significantly better than precooling (P < 0.05). Counter-irritation was significantly more effective than precooling methods in reduction of pain perception and improvement of behavioral feedback (P < 0.05). Conclusions:The perceived pain was significantly lower when the benzocaine method was used. Behavioral feedback was significantly better with benzocaine and counter-irritation methods compared to precooling procedures.


A study on prophagic and chromosomal sodC genes involvement in Escherichia coli O157:H7 biofilm formation and biofilm resistance to H2O2.
Escherichia coli O157:H7 possesses one chromosomal and two prophagic sodC genes encoding for Cu,Zn superoxide dismutases. We evaluated the contribution of sodC genes in biofilm formation and its resistance to hydrogen peroxide. The biofilm of sodC deletion mutants has been studied, in presence or absence of hydrogen peroxide, by crystal violet in 96-well plates and Scanning Electron Microscopy on glass coverslips. Deletion of prophagic sodC genes had no effect on biofilm construction, in contrast to the chromosomal gene deletion. Hydrogen peroxide treatment showed higher cell mortality and morphological alterations in sodC deletion mutants respect to wild type. These effects were related to the biofilm development stage. The role of the three SodCs is not redundant in biofilm formation and the resistance to oxidative damage. The stage of biofilm development is a crucial factor for an effective sanitization.


Lipopolysaccharide, a key molecule involved in the synergism between temporins in inhibiting bacterial growth and in endotoxin neutralization.
Lipopolysaccharide (LPS) is the major structural component of the outer membrane of Gram-negative bacteria and shields them from a variety of host defense factors, including antimicrobial peptides (AMPs). LPS is also recognized by immune cells as a pathogen-associated molecular pattern and stimulates them to secrete pro-inflammatory cytokines that, in extreme cases, lead to a harmful host response known as septic shock. Previous studies have revealed that a few isoforms of the AMP temporin, produced within the same frog specimen, can synergize to overcome bacterial resistance imposed by the physical barrier of LPS. Here we found that temporins can synergize in neutralizing the LPS-induced macrophage activation. Furthermore, the synergism between temporins, to overcome the protective function of LPS as well as its endotoxic effect, depends on the length of the polysaccharide chain of LPS. Importantly, mode of action studies, using spectroscopic and thermodynamic methods, have pointed out different mechanisms underlying the synergism of temporins in antimicrobial and anti-endotoxin activities. To the best of our knowledge, such a dual synergism between isoforms of AMPs from the same species has not been observed before, and it might explain the ability of such amphibians to resist a large repertoire of microorganisms.


Rapid and sensitive quantification of Borrelia burgdorferi-infected mouse tissues by continuous fluorescent monitoring of PCR.
The quantity of Borrelia burgdorferi organisms in tissue samples is an important determinant for infection studies in the mouse model of Lyme disease. This report presents the development of a rapid and sensitive external-standard-based PCR assay for the absolute quantification of B. burgdorferi in mouse tissue samples. The assay uses a double-stranded DNA dye to continuously monitor product formation and in less than an hour was able to quantify samples ranging up to 6 log units in concentration. The PCR efficiencies of the sample and the standard were matched by using a standard composed of purified B. burgdorferi chromosome mixed with tissue-matched mouse genome lacking bacterial DNA. Normalization of B. burgdorferi quantities to the mouse nidogen gene allowed comparison of B. burgdorferi numbers in samples isolated from different tissues and strains. PCR analysis of the chromosomal gene recA in cultured B. burgdorferi was consistent with a single recA per bacterium. The parameters defined in this assay should be applicable to quantification of other organisms, even infectious agents for which no ready source of DNA standard is available. In summary, this report presents a rapid external-standard-based PCR method for the quantification of B. burgdorferi in mouse DNA samples.