Clinical and laboratory outcomes during the treatment of rheumatoid arthritis with methotrexate.
Ten clinical and three laboratory outcomes were evaluated in 86 patients completing a double-blind placebo-controlled trial of methotrexate in rheumatoid arthritis. The improvement in all measured outcomes was statistically significantly better in patients receiving methotrexate than in patients receiving placebo. The correlations of the changes in outcome measures were calculated to determine if improvement in one parameter was associated with improvement in other clinical parameters. Associations between different clinical outcomes were often statistically significant. Associations between laboratory outcomes were also often statistically significant. However, the association between clinical outcomes and laboratory outcomes was generally poor.


Group 3 LEA Gene HVA1 Regulation by Cold Acclimation and Deacclimation in Two Barley Cultivars with Varying Freeze Resistance.
The level of expression of the group 3 late embryogenesis abundant abscisic acid-regulated gene (HVA1) to cold treatment has been studied in winter barley (Hordeum vulgare) seedling tissue. The cDNA clone (pHVA1) encoding this late embryogenesis abundant protein was used as a hybridization probe to detect the corresponding mRNA. Expression of the HVA1 gene was determined after the tissue had been subjected to a regimen of 2 degrees C exposure (cold acclimation), followed by a return to 25 degrees C growth conditions (deacclimation). Accumulation of HVA1 mRNA occurred upon cold acclimation of the tissue and disappeared as early as 2 hours after exposure to deacclimation conditions. A comparison of the response to cold acclimation and deacclimation was made between seedling tissue of a freeze-resistant and less freeze-resistant cultivar. In both cultivars, the HVA1 gene was expressed and modulated by cold treatment. Within 2 hours of deacclimation HVA1 mRNA was no longer detectable in either cultivar independently of freeze resistance. The level of expression of HVA1 appeared to be greater in the less freeze-resistant cultivar (Winter Malt).


Necrotizing granulomatous inflammation: what does it mean if your special stains are negative?
Necrotizing granulomas are commonly encountered in surgically resected specimens. The majority will be proven infectious with special stains for microorganisms. These need to be distinguished from other granulomatous processes such as Wegener's granulomatosis (WG). Although there may be histological overlap between these different processes, the identification of a true necrotizing vasculitis is specific to WG in the context of necrotizing granulomas. Otherwise, the combination of histological features should lead to a specific diagnosis. Despite a thorough histological examination and assessments of special stains, a significant proportion of necrotizing granulomas will appear infectious with no obvious infectious etiology. There are only few clinically available ancillary tests that can be performed on paraffin-embedded tissue and include real-time PCR for tuberculous mycobacteria. Despite correlation with clinical, serological and other microbiological studies, some necrotizing granulomas remain unexplained. Patients with such granulomas appear not to require any additional treatment and do experience a favorable outcome.


Differential storage of prolactin, granins (Chromogranin B and secretogranin II), and constitutive secretory markers in rat pituitary GH4C1 cells.
The rat pituitary cell line GH4C1 secretes granins (chromogranin B and secretogranin II) and prolactin by the regulated secretory pathway. The intracellular storage of prolactin is preferentially induced by hormone treatment with estradiol, insulin, and epidermal growth factor. The goal of this study was to determine the effect of hormone treatment on storage of granins and constitutive secretory markers. The granins were efficiently stored in both hormone-treated and -untreated cells (17% of total secreted in 4 h). Secreted alkaline phosphatase (SEAP), a truncated membrane protein that would not be expected to enter secretory granules, and glycosaminoglycan, a marker for the constitutive secretory pathway, exhibited 70 80% secretion under both conditions. In comparison, the relative prolactin secretion was 31 and 68% from hormone-treated and -untreated cells, respectively. Phorbol ester and KCl stimulated prolactin secretion 2.3-fold from untreated cells and 5. 5-fold from hormone-treated cells. In contrast, SEAP secretion was stimulated 1.5-fold from both treated and untreated cells, consistent with secretion by the constitutive secretory pathway. Stimulated secretion of granins was detected from both hormone-treated and -untreated cells. These results suggest that granin and prolactin storage are differentially regulated and that the constitutive secretory pathway is not affected by hormone treatment.


Refractory chronic pain.
Refractory chronic pain is a significant public health problem and frustrating to everyone affected by it. All physicians can participate in the care of these patients, but psychiatrists should take a leading role in their care. A comprehensive approach offers hope and increases the opportunities for successful treatment. Each perspective of an interdisciplinary formulation has a unique logic that defines specific methods for designing treatment for the patient with refractory chronic pain. The patient does not have to fit into one theoretic approach or model to receive and accept treatment. The patient's diagnoses are based on the formulation, which then directs treatment along rational directions. The linkages and interactions of a patient's diagnoses can then be investigated within a framework that includes the entire person and not just his or her biochemistry. If a patient's suffering persists, other factors must be considered that may have been overlooked before the treatment plan is abandoned or modified. Usually these factors are within one of the perspectives initially thought to be less important. A new combination of approaches is then required to treat the patient successfully. The perspectives appreciate not only that the patient is struggling through important life events but also that he is a person composed of vulnerabilities and strengths, having made many choices and afflicted by diseases.


Predictive value of P-selectin and endothelin-1 for vascular restenosis after interventional procedures for peripheral artery disease.
Peripheral artery disease (PAD) is caused by the building up of plaques in the arteries that carry blood to the lower limbs. The present study aimed to assess the predictive value of the plasma levels of P-selectin (Ps) or endothelin-1 (ET-1) regarding the occurrence of vascular restenosis after endovascular therapy for PAD. Patients with or without vascular restenosis confirmed by computed tomography angiography after endovascular therapy between March and December 2015 (n=20 per group) were enrolled. The serum levels of Ps and ET-1 prior to the operation and at 1 h, as well as 1, 2 and 3 weeks after the operation were compared between the two groups. At 1 h after the operation, the serum levels of Ps and ET-1 were significantly increased as compared with the pre-operative levels (P<0.05). The serum levels of Ps and ET-1 at 1 h, as well as 1, 2 and 3 weeks after the operation in the restenosis group were significantly higher as compared with those in the non-stenosis group (P<0.05). However, for the non-stenosis group, the serum levels of Ps and ET-1 at 1, 2 and 3 weeks after the operation did not significantly differ from the pre-operative levels (P>0.05). The diagnostic sensitivity and specificity of the serum ET-1 levels at 1 h after the operation for predicting post-operative restenosis in PAD patients with a cut-off of 0.1089 pg/ml were 85 and 85%, respectively. In conclusion, the serum levels of Ps and ET-1 have a high predictive value for post-operative vascular restenosis after endovascular therapy for PAD patients.


Urinary neutrophil gelatinase-associated lipocalin: a marker of urinary tract infection among febrile children.
Neutrophil gelatinase-associated lipocalin (NGAL) has emerged as a valuable biomarker of urinary tract infection (UTI) in children. This study aimed to compare the diagnostic accuracy of urinary NGAL (uNGAL) with those of serum C-reactive protein (CRP) and white blood cell (WBC) count for predicting UTI and acute pyelonephritis (APN) in febrile children. The medical charts of children undergoing uNGAL measurements between November 2017 and August 2019 were retrospectively reviewed. Patients with a suspected or diagnosed UTIs were included. The diagnostic accuracies of uNGAL, serum CRP, and WBC count for detecting UTI and APN were investigated. Independent predictors of UTI and APN were investigated using multivariable logistic regression analyses. A total of 321 children were enrolled in this study. The uNGAL levels were higher in the UTI group (n=157) than in the non-UTI group (n=164) (P<0.05). Among children with a UTI, uNGAL levels were higher in the APN group (n=70) than, the non-APN group (n=87) (P<0.05). In the multivariate analysis, uNGAL was independently associated with UTI and APN (both P<0.05). Serum CRP and WBC count were not correlated with the presence of UTI and APN. Receiver operating curve analyses showed that the uNGAL level had the highest area under the curve (AUC) for predicting UTI and APN, respectively (AUC, uNGAL vs. CRP vs. WBC count, 0.860 vs. 0.608 vs. 0.669 for UTI; 0.780 vs. 0.680 vs. 0.639 for APN, all P<0.05, respectively). The predictive values and likelihood ratios of uNGAL were superior to those of serum CRP and WBC count for detecting UTI and APN at each cutoff level. UNGAL may be more useful than serum CRP and WBC count for identifying and assessing UTI in febrile children.


Acute Acquired Methemoglobinemia Following Nitrobenzene Poisoning.
Nitrobenzene is an aromatic compound commonly used in agricultural fertilizers. It is capable of inducing methemoglobinemia when ingested in sufficient quantities. Methemoglobinemia impairs oxygen transport by dual mechanism of impaired oxygen binding and diminished oxygen unloading. We report a case of nitrobenzene poisoning accompanied by methemoglobinemia and later complicated by unconjugated hyperbilirubinemia secondary to hemolysis. Prompt treatment with intravenous methylene blue based on a clinical diagnosis in view of dropping SpO2 unresponsive to oxygen with chocolate cyanosis and paradoxically elevated PaO2 enabled favorable patient outcome. Other recommended modalities of treatment in acquired acute methemoglobinemia include vitamin C, hyperbaric oxygen and exchange transfusions. Keywords: Methemoglobinemia; methylene blue; nitrobenzene; nitroboom.


[Endoscopic treatment of vesicoureteral reflux].
Endoscopic subureteral injection of tissue-augmenting substances, a 15-minute outpatient procedure has become an alternative to long-term antibiotic prophylaxis and surgical intervention in the treatment of vesicoureteral reflux (VUR) in children. We searched MEDLINE using the words: vesicoureteral reflux, treatment, the long-term results of endoscopic treatment of reflux. We summarized the worldwide data regarding endoscopic treatment of VUR using various tissue-augmenting substances presently available. In terms of effectiveness and long-term successful results, polytetrafluoroethylene is still the most reliable injectable material for the endoscopic treatment of VUR. However, Dextranomer/hyaluronic acid copolymer (Deflux) is a new promising tissue augmenting substance which might be able to replace Teflon in the endoscopic treatment of reflux in terms of a similar to Teflon reflux cessation rate and exhibiting no evidence of migration. Endoscopic subureteral polytetrafluoroethylene injection is a simple and effective outpatient procedure for in the treatment of vesicoureteral reflux. No long-term morbidity was observed in our patients with small amounts of injectable polytetrafluoroethylene.


[The effect of maternal education on the risk of infant mortality].
The impact of maternal education on infant mortality in Brazil is analyzed by estimating relative risks in a case control study. "The cases consisted of a sample of infant deaths which were registered at the Registry Office and which occurred in the Metropolitan Region of Porto Alegre (Brazil) in 1980, and the control group was a sample of births occurring in the same region in 1980, and also notified to the Registry Office. With information obtained from a special household survey, a total of 688 cases and 762 controls [were] analyzed...for maternal education and its association with family income, birth order and environmental factors. Specific relative risks (associated with maternal education) have also been calculated separately for 3 groups of cause-of-death and for 3 age-at-death groups." (SUMMARY IN ENG)


Bacteriologic evaluation of the effect of Nd:YAG laser irradiation in experimental infected root canals.
The aim of this study was to evaluate the efficacy of the Pumped Diodium-Nd:YAG laser in sterilizing contaminated root canals. After hand instrumentation, 30 teeth were inoculated with Actinomyces naeslundii CH-12 and 30 teeth with Pseudomonas aeruginosa ATCC 27853 and incubated for 24 h. The teeth were divided into three subgroups: subgroup A received no treatment; subgroup B was irradiated with laser (5 Hz for 15 s or 10 Hz for 15 s); and subgroup C was irrigated with 5.25% NaOCl. The number of viable bacteria in each group was evaluated by using the surface-spread plate technique. The results indicated an average of 34.0% decrease in colony-forming units for A. naeslundii CH-12 and 15.7% for P. aeruginosa ATCC 27853 with the 5 Hz/15 s laser treatment, and for the 10 Hz laser frequency, a decrease of the 77.4% for A. naeslundii CH-12 and 85.8% for P. aeruginosa ATCC 27853. No bacteria were detected in the canals treated with 5.25% NaOCl. The results show an antibacterial effect of the Pumped Diodium Nd:YAG laser, depending on the radiation frequency. However, 5.25% NaOCl was more effective than either laser application.


Treatment cost for typhoid fever at two hospitals in Kolkata, India.
The purpose of this study was to estimate treatment cost for typhoid fever at two hospitals in Kolkata, India. This study was an incidence-based cost-of-illness analysis from the providers' perspective. Micro-costing approach was employed for calculating patient-specific data. Unit costs of medical services used in the calculation were directly measured from the study hospital by standard method. The study hospitals were selected based on accessibility to data and cooperation. Eighty-three Widal-positive and/or culture-confirmed patients with typhoid fever during November 2003-April 2006 were included in the study. Most (93%) patients were children. Eighty-one percent was treated at the outpatient department. The average duration of hospitalization for child and adult patients was 8.4 and 4.2 days respectively. The average cost of treating children, adults, and all patients was US$ 16.72, 72.71, and 20.77 respectively (in 2004 prices). Recalculation based on 80% occupancy rate in inpatient wards (following the recommendation of the World Health Organization) found that the cost of treating children, adults, and all patients was US$ 14.53, 36.44, and 16.11 respectively.


Spectroscopic identification of different types of copper centers generated in synthetic four-helix bundle proteins.
Using a combined rational-combinatorial approach, stable copper binding sites were implemented in template-assembled synthetic four-helix bundle proteins constructed by three different helices with only 16 amino acid residues. These peptides include two histidines and one cysteine at positions appropriate for coordinating a copper ion. Sequence variations of the helices were made in the second coordination shell or even more remote from the copper binding site (i) to increase the overall stability of the metalloproteins and (ii) to fine-tune the structure and properties of the copper center. As a result, ca. 90% of the 180 proteins that were synthesized were capable to bind copper with a substantially higher specificity than those obtained in the first design cycle (Schnepf, R.; Horth, P.; Bill, E.; Wieghardt, K.; Hildebrandt, P.; Haehnel, W. J. Am. Chem. Soc. 2001, 123, 2186-2195). Furthermore, the stabilities of the copper protein complexes were increased by up to 2 orders of magnitude and thus allowed a UV-vis absorption, resonance Raman, electron paramagnetic resonance, and (magnetic) circular dichroism spectroscopic identification and characterization of three different types of copper binding sites. It could be shown that particularly steric perturbations in the vicinity of the His(2)Cys ligand set control the formation of either a tetragonal (type II) or a tetrahedral (type I) copper binding site. With the introduction of two methionine residues above the histidine ligands, a mixed-valent dinuclear copper binding site was generated with spectroscopic properties that are very similar to those of Cu(A) sites in natural proteins. The results of the present study demonstrate for the first time that structurally different metal binding sites can be formed and stabilized in four-helix bundle proteins.


Multiple intracranial tuberculomas in a child.
Intracranial tuberculomas continue to be a serious complication of central nervous system tuberculosis. Tuberculomas are conglomerates of tubercles resulting from hematogenous spread of infection. Modern neuroimaging studies such as magnetic resonance imaging and molecular biologic techniques such as polymerase chain reaction are helpful in the diagnosis of central nervous system tuberculosis and tuberculomas. We report a boy with multiple intracranial tuberculomas diagnosed and treated with the aids of magnetic resonance and polymerase chain reaction techniques.


Nurse-administered propofol sedation: feasibility and safety in bronchoscopy.
Propofol is a fast-acting intravenous sedative that has advantages as a procedural sedative over traditional regimens. It has been shown to have a similar safety profile to traditional sedating medications in the setting of gastroenterologic endoscopy. Nurse-administered propofol sedation is given by a specially-trained nurse, without anesthesiologist involvement. We have used nurse-administered propofol sedation in our bronchoscopy suite for several years. In this report, we summarize our experience with nurse-administered propofol sedation, and demonstrate it to be feasible and safe for bronchoscopic procedures. Procedure reports and nursing notes for 588 bronchoscopic procedures performed between July 2006 and June 2008 were retrospectively reviewed. Patient demographics, procedure type and indication, procedure time, medication doses, and adverse events were noted and analyzed. Nurse-administered propofol sedation was used in 498/588 (85%) procedures. Patients utilizing nurse-administered propofol sedation had an average age of 53 years (range 18-86) with an average weight of 80 kg. 56% of the patients were male, and 57% of the procedures were performed on outpatients. Average procedure duration was 25 min (range 3-123). The average propofol dose was 3.13 mg/kg (range 0.12-20 mg/kg). Adverse events attributable to sedation were noted in 33 (6.6%) procedures. Of the 14 (2.8%) major adverse events (death, need for intubation, ICU stay, or hospitalization), only 6 (1.2%) were potentially attributable to the sedation regimen. There were 2 deaths, neither of which was related to sedation. Nurse-administered propofol sedation is a feasible and safe sedation method for bronchoscopic procedures.


Growth characteristics of enterococci isolated from Laban Rayeb.
Eleven strains of Enterococcus (Streptococcus) faecalis isolated from Laban Rayeb, a type of fermented milk, had been studied when grew in milk. Enterococcus (Streptococcus) faecalis strains 19 and 22 were the most active strains in the production of lactic acid, acetaldehyde and diacetyl. Both strains exhibited a high proteolytic activity besides a good clean flavour. Based on the obtained results, Enterococcus (Streptococcus) faecalis 19 and 22 could be used to bring about the lactic fermentation during the manufacture of some cultured dairy products.


Accuracy of Magnetic Resonance Imaging for Identifying Ovarian Cancer in a Community-Based Setting.
Introduction: Many ovarian or adnexal masses have an indeterminate appearance on ultrasound that can raise concerns about cancer. Although magnetic resonance imaging (MRI) has been reported to reliably distinguish between benign and malignant masses, studies evaluating the accuracy of MRI in community-based practice settings are lacking. Methods: Women who underwent MRI to further evaluate an ultrasound-detected adnexal mass in 2016-2017 within a large community-based health system were identified. MRI reports were classified as favoring malignancy, benign disease, or indeterminate, blinded to pathological outcome. With a minimum of 2 years of follow-up, all ovarian cancers and borderline tumors were identified, and the accuracy of MRI assessment was determined. Results: Among 338 women who had MRI to evaluate an adnexal mass, 144 (42.6%) subsequently underwent surgery. MRI favored malignancy in 7 (4.9%) cases, benign disease in 89 (62.2%) cases, and was indeterminate in 48 (33.6%) cases. Of the seven cases in which MRI favored malignancy, two cancers and five benign tumors were found. An additional 10 cases of cancer or borderline tumor were found among women who had MRI reports that were read as indeterminate (n = 6) or that favored benign disease (n = 4). The sensitivity, specificity, positive predictive value, and negative predictive value of an MRI favoring malignancy were 16.7%, 96.2%, 28.5%, and 92.7%, respectively. Discussion: In a large community-based setting, an MRI favoring malignancy was more likely to be associated with benign disease than cancer and identified only 16.7% of true malignant cases. The findings suggest that the ability of MRI to differentiate between benign and malignant adnexal masses in community-based practice settings is currently limited.


Ischemia/Reperfusion Injury of Fatty Liver Is Protected by A2AR and Exacerbated by A1R Stimulation through Opposite Effects on ASK1 Activation.
Hepatic ischemia/reperfusion injury (IRI) is aggravated by steatosis and is a main risk factor in fatty liver transplantation. Adenosine receptors (ARs) are emerging as therapeutic targets in liver diseases. By using cellular and in vivo systems of hepatic steatosis and IRI, here we evaluated the effects of pharmacological A2AR and A1R activation. The A2AR agonist CGS21680 protected the primary steatotic murine hepatocyte from IR damage and the activation of ASK1 and JNK. Such an effect was attributed to a phosphatidylinositol-3-kinase (PI3K)/Akt-dependent inhibition of ASK1. By contrast, the A1R agonist CCPA enhanced IR damage, intracellular steatosis and oxidative species (OS) production, thereby further increasing the lipid/OS-dependent ASK1-JNK stimulation. The CGS2680 and CCPA effects were nullified by a genetic ASK1 downregulation in steatotic hepatoma C1C7 cells. In steatotic mice livers, CGS21680 protected against hepatic IRI and ASK1/JNK activation whereas CCPA aggravated hepatic steatosis and IRI, and enhanced ASK1 and JNK stimulation. These results evidence a novel mechanism of CGS21680-mediated hepatoprotection, i.e., the PI3K/AKT-dependent inhibition of ASK1, and they show that CGS21680 and CCPA reduces and enhances the IRI of fatty liver, respectively, by preventing or increasing the activation of the cytotoxic ASK1/JNK axis. They also indicate the selective employment of A2AR agonists as an effective therapeutic strategy to prevent IRI in human fatty liver surgery.


Generalized fluctuation-dissipation and reciprocal relations for Brownian sieves and molecular machines.
A Brownian sieve is a spatially periodic microstructured device that combines the effects of thermal noise, local spatial asymmetry, and external forces to separate particles based on their transport properties. By treating the motion of an individual particle as a cyclical process in which the particle fluctuates away from and returns to the origin of some unit cell, I derive generalized fluctuation-dissipation and reciprocal relations for the averages (and for all moments) of the number of periodic displacements that are exact and valid for arbitrary values of the external forces. These relations hold not only for Brownian sieves, but for all molecular machines in which a nanoscale system couples two chemical, mechanical, or transport processes by a cycle in which the molecular machine itself fluctuates away from and then returns to some arbitrary reference state, in the process doing or receiving work on or from the environment.


Persistence of Norovirus GII Genome in Drinking Water and Wastewater at Different Temperatures.
Human norovirus (NoV) causes waterborne outbreaks worldwide suggesting their ability to persist and survive for extended periods in the environment. The objective of this study was to determine the persistence of the NoV GII genome in drinking water and wastewater at three different temperatures (3 °C, 21 °C, and 36 °C). The persistence of two NoV GII inoculums (extracted from stool) and an indigenous NoV GII were studied. The samples were collected for up to one year from drinking water and for up to 140 days from wastewater. Molecular methods (RT-qPCR) were used to assess the decay of the NoV genome. Decay rate coefficients were determined from the fitted decay curves using log-linear and/or non-linear model equations. Results showed significant differences in the decay kinetics of NoV genome between the temperatures, matrices, and virus strains. The persistence of NoV was higher in drinking water compared to wastewater, and the cold temperature assisted persistence at both matrices. Differences between the persistence of NoV strains were also evident and, particularly, indigenous NoVs persisted better than spiked NoVs in wastewater. The decay constants obtained in this study can be utilized to assess the fate of the NoV genome in different water environments.


Surveillance studies involving HIV testing are needed: Will at-risk youth participate?
Adolescent males who have sex with males (AMSMs) account for high numbers of new HIV diagnoses. To date, surveillance data have been limited to diagnosed cases of HIV, resulting in an underestimation of risk and burden among AMSMs unwilling or unable to access HIV testing. This study identified facilitators and barriers to AMSMs' participation in future surveillance studies involving HIV testing. AMSMs (n = 198) aged 14 to 17 years participated. The majority identified as non-Hispanic White or Latinx, had a least 1 male sex partner, and self-reported HIV negative. Participants read an online survey beginning with a vignette describing a hypothetical HIV surveillance study requiring HIV testing. They then completed questions assessing likelihood to participate, perceived research benefits and risks, attitudes toward HIV risk, prior HIV health services, and parental awareness of sexual orientation. Approximately 40% indicated strong willingness to participate. Willingness was positively related to perceived HIV risk, free access to HIV testing, counseling and referral if testing positive, confidentiality protections, and lack of access to a trusted physician. Having to tell others if one tested positive for HIV and requirements for guardian permission were significant participation barriers. Inclusion of HIV testing in surveillance studies is essential for accurate estimation of HIV incidence and prevalence among AMSMs. Successful recruitment of sexual minority youth into sexual health surveillance research will require procedures tailored to youth's health care needs and concerns, including adequate HIV counseling, referral to treatment if seropositive, and attention to concerns regarding guardian permission. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Is there concordance between overactive bladder and detrusor overactivity in men with predominant storage urinary symptoms referred to Functional Urology and Urodynamics Units?
To assess the diagnostic concordance of overactive bladder (OAB) and detrusor overactivity (DO) in male patients with predominant storage lower urinary tract symptoms (LUTS) and evaluate their clinical and urodynamic profile according to DO presence and degree of obstruction. Epidemiological, cross-sectional multicenter study. A 3-day bladder diary (3dBD), International Prostate Symptom Score (IPSS) and Bladder Control Self-Assessment Questionnaire (B-SAQ) questionnaires were analyzed. Prostate volume was determined by ultrasound. Urodynamic study (UDS) tests were performed. The prevalence of OAB and DO and the degree of clinical concordance (kappa index) were investigated. Descriptive analysis of clinical variables and UDS results was performed, followed by comparisons based on the presence of DO and degree of obstruction. A total of 445 patients were included. The mean age was (SD) 54.8 (9.9) years. According to 3dBD, 89.9% presented increased urinary frequency, 87.9% nocturia, 72.1% urgency, and 31.9% urge urinary incontinence (UUI). Obstruction was present in 36.8%. Concomitant OAB and DO were present in 54.5%. The degree of diagnostic concordance between OAB and DO was low (κ = 0.1772). There were more patients with DO presenting urgency (3dBD and B-SAQ; p < 0.001), UUI (3dBD; p = 0.008) and nocturia (B-SAQ; p < 0.001). Differences were found in terms of prostate volume, IPSS-voiding, maximum flow (Qmax) and post-void residual (p < 0.05) according to the obstruction degree. Approximately 50% of male patients aged 18-65 years old with predominant storage LUTS, referred to specialized units, have both OAB and DO. Obstruction is present on 1/3. Diagnostic concordance between OAB and DO is poor.


Meiotic segregation analysis of RB1 alleles in retinoblastoma pedigrees by use of single-sperm typing.
In hereditary retinoblastoma, different epidemiological studies have indicated a preferential paternal transmission of mutant retinoblastoma alleles to offspring, suggesting the occurrence of a meiotic drive. To investigate this mechanism, we analyzed sperm samples from six individuals from five unrelated families affected with hereditary retinoblastoma. Single-sperm typing techniques were performed for each sample by study of two informative short tandem repeats located either in or close to the retinoblastoma gene (RB1). The segregation probability of mutant RB1 alleles in sperm samples was assessed by use of the SPERMSEG program, which includes experimental parameters, recombination fractions between the markers, and segregation parameters. A total of 2,952 single sperm from the six donors were analyzed. We detected a significant segregation distortion in the data as a whole (P=.0099) and a significant heterogeneity in the segregation rate across donors (.0092). Further analysis shows that this result can be explained by segregation distortion in favor of the normal allele in one donor only and that it does not provide evidence of a significant segregation distortion in the other donors. The segregation distortion favoring the mutant RB1 allele does not seem to occur during spermatogenesis, and, thus, meiotic drive may result either from various mechanisms, including a fertilization advantage or a better mobility in sperm bearing a mutant RB1 gene, or from the existence of a defectively imprinted gene located on the human X chromosome.


An Evaluation of Contraception Education and Health Promotion Applications for Patients.
Patients use mobile applications (apps) to obtain information about health, including contraception. Providers and health educators may also use apps designed for patients to aid in patient education during the clinical encounter or recommend apps for patient use. Individuals may have a difficult time remaining updated on the number and quality of available apps. The objective of this study is to identify and evaluate contraception apps for patient education and health promotion, so that providers and health educators may recommend accurate apps to patients. We systematically searched the Apple iTunes Store using contraception search terms. A master list of apps was created and the apps were divided into categories and subcategories according to intended audience and purpose. Contraception apps for patient education and health promotion were selected and also checked for availability in the Google Play Store. We evaluated these identified apps using an adapted APPLICATIONS scoring system. Forty-eight apps were identified from the original search. Nineteen of these were excluded because they did not open on an iPhone or iPad, were no longer available, or did not contain educational material on contraception. We excluded 11 additional apps that contained inaccurate information. We evaluated 18 apps. The mean score was 10.6 out of 17 possible points with a range of 7 to 15 points. Many apps provide contraception information for patients, but some apps are inaccurate. Few apps provide comprehensive information on all available methods, including effectiveness, side effects, and contraindications.


[Indicators for ambulatory quality management in patients with bronchial asthma].
The process of quality assurance and quality improvement in ambulatory care is bound to the development of quality indicators. The performance and outcomes of medical services offered by physicians have to be assessed on the basis of valid, reliable and practicable indicators. The aim of this analysis was to identify relevant process and outcome measures which could be used as quality indicators in ambulatory care of asthmatic patients in Germany. Therefore, the process of routine ambulatory care of asthmatic patients was analyzed and compared to existing management guidelines of national and international expert panels. A set of 51 indicators was derived from this analysis and a systematic review of 96 articles on quality management of asthma care selected after research in MEDLINE 1966-1997. This list of quality indicators should be discussed in quality circles for practicability in ambulatory care settings.


Activation of the mitogen-activated protein kinase p38 by human cytomegalovirus infection through two distinct pathways: a novel mechanism for activation of p38.
Recent evidence indicates activated mitogen-activated protein kinase (MAPK) p38 has a critical function in human cytomegalovirus (HCMV) viral DNA replication in infected human fibroblasts. To elucidate the mechanism of HCMV-mediated p38 activation, we have performed a detailed analysis of p38 activation and the kinases associated with this activation at different times postinfection. We demonstrate that p38 kinase activity is strongly increased following viral infection. Inhibition of this activity significantly inhibited HCMV-induced hyperphosphorylation of pRb and phosphorylation of heat shock protein 27, suggesting that p38 activation is involved in virus-mediated changes in host cell metabolism throughout the course of infection. We then provide evidence that p38 activation is mediated by different mechanisms at early times versus later times of infection. At early times of infection (8 to 14 h postinfection [hpi]), when p38 activation is first observed, no significant activation of the three kinases which can directly phosphorylate p38 (namely, MKK3, MKK6, and MKK4) is detected. Using vectors which express dominant negative proteins, we demonstrate that basal MKK6 kinase activity is necessary for HCMV-mediated p38 activation at these early times of infection (12 hpi). Then, we use ATP depletion to show that at 12 hpi, HCMV inhibits dephosphorylation of activated p38. These two experiments suggest that HCMV activates p38 by inhibition of dephosphorylation of p38. In contrast to early times of infection, at later times of infection (48 to 72 hpi), increased MKK3/6, but not MKK4, activity is observed. These results indicate that at early times of HCMV infection, increased steady-state levels of activated p38 is mediated at least in part by inhibition of dephosphorylation of p38, while at later times of infection p38 activation is due to increased activity of the upstream kinases MKK3 and MKK6. These findings indicate that HCMV has developed multiple mechanisms to ensure activation of the MAPK p38, a kinase critical to viral infection.


Missed diagnosis of POEMS syndrome with onset of progressive fatigue and numbness: a case report.
POEMS syndrome is a rare multisystem disease associated with an underlying plasma cell disorder. Its name is an acronym for peripheral neuropathy (P), endocrinopathy (E), organomegaly (O), monoclonal plasma cell proliferative disorder (M), and skin changes (S). This case report describes a patient with POEMS syndrome who presented with progressive fatigue and numbness in the lower extremities. Initially, the patient was erroneously diagnosed with diabetes and diabetic peripheral neuropathy because of the endocrinopathy associated with POEMS syndrome. After a second hospitalization, the patient was diagnosed with POEMS syndrome and recovered with alkylator therapy and a peripheral blood stem cell transplant. The patient's overall condition was improved at the 1-year follow-up. POEMS syndrome should be considered if a patient presents with endocrinopathy and unexplained peripheral neuropathy.


Dental erosion: diagnostic-based noninvasive treatment.
Lesions that result from dental erosion can be difficult to recognize--particularly when abrasion and attrition are also present. Consequently, dental erosion is often misdiagnosed and mistreated by radical restorative modalities that compromise the vitality of the pulp. This article provides clinicians with knowledge concerning the diagnosis of the complex lesions of dental wear and demonstrates the conservative treatment of this condition. Two cases that exhibit marked tooth wear in anterior teeth and their subsequent restoration utilizing occlusal principles and composite resin are presented.


Background matters: Minor vibratory stimulation during motor skill acquisition selectively reduces off-line memory consolidation.
Although a ubiquitous situation, it is not clear how effective is a learning experience when task-irrelevant, sensory noise occurs in the background. Here, young adults were trained on the finger opposition sequence task, in a well-established training and testing protocol affording measures for online as well as off-line learning. During the training session, one group experienced a minor background vibratory stimulation to the trunk by the means of vibrating cushion, while the second group experienced recorded sound vibrations. A control group was trained with no extra sensory stimulation. Sensory stimulation during training had no effect on the online within-session gains, but dampened the expression of the off-line, consolidation phase, gains in the two sensory stimulation groups. These results suggest that background sensory stimulation can selectively modify off-line, procedural memory consolidation processes, despite well-preserved on-line learning. Classical studies have shown that neural plasticity in sensory systems is modulated by motor input. The current results extend this notion and suggest that some types of task-irrelevant sensory stimulation, concurrent with motor training, may constitute a 'gating' factor - modulating the triggering of long-term procedural memory consolidation processes. Thus, vibratory stimulation may be considered as a behavioral counterpart of pharmacological interventions that do not interfere with short term neural plasticity but block long-term plasticity.


The Emerging Role of the Gut Microbiome in Adult Patients With Multiple Sclerosis.
Approximately 2.3 million people worldwide are currently living with multiple sclerosis (MS). The pathophysiologic mechanism of MS is not well known. It has been suggested that alterations in the normal gut flora may contribute to MS etiology and symptoms. The aims of this review are to describe the data suggesting a role for the gut microbiome in MS research and address its implications for practice. A literature search of the following databases (PubMed, CINAHL, Cochrane library database, MEDLINE, Scopus, and Psychology and Behavioral Sciences) was conducted to find published studies relevant to gut microbiome in patients with MS. Five articles met the inclusion criteria of research studies of human gut microbiome in adults in English language and those receiving disease-modifying medications. Exclusion criteria were case reports and reviews. Human studies found that the gut microbiome was different among patients with MS, patients with MS who were treated with glatiramer acetate, and healthy controls. There is beginning evidence to suggest that the gut microbiota is related to autoimmunity and the pathology of MS. However, more research is necessary to clarify these mechanisms. A better understanding of the role of the gut microbiota in MS may lead to the development of targeted individualized interventions affecting the gut microbiota. These interventions may emphasize symptom self-management strategies such as diet.


Succession of bacterial community structure along the Changjiang River determined by denaturing gradient gel electrophoresis and clone library analysis.
Bacterial community structure along the Changjiang River (which is more than 2,500 km long) was studied by using denaturing gradient gel electrophoresis (DGGE) and clone library analysis of PCR-amplified 16S ribosomal DNA (rDNA) with universal bacterial primer sets. DGGE profiles and principal-component analysis (PCA) demonstrated that the bacterial community gradually changed from upstream to downstream in both 1998 and 1999. Bacterial diversity, as determined by the Shannon index (H'), gradually decreased from upstream to downstream. The PCA plots revealed that the differences in the bacterial communities among riverine stations were not appreciable compared with the differences in two adjacent lakes, Lake Dongting and Lake Poyang. The relative stability of the bacterial communities at the riverine stations was probably due to the buffering action of the large amount of water flowing down the river. Clone library analysis of 16S rDNA revealed that the dominant bacterial groups changed from beta-proteobacteria and the Cytophaga-Flexibacter-Bacteroides group upstream to high-G+C-content gram-positive bacteria downstream and also that the bacterial community structure differed among the stations in the river and the lakes. The results obtained in this study should provide a reference for future changes caused by construction of the Three Gorges Dam.


Malignant tumors in an ancient Egyptian population.
Since it is still an open debate whether malignant tumors are mainly influenced by environmental factors, the frequency of such malignant tumors in historic populations with different living conditions is of particular interest. In the present study, we investigated the occurrence of malignant tumors affecting bone tissue in a population of mumrnies and skeletons, which had been excavated from the large necropolis of Thebes-West, Upper Egypt. Our study material comprised a series of at least 415 individuals (thereof 325 adults) dating from approx. 1500-500 B.C. All individuals had been mummified, but were severely damaged and partially broken by grave robbers, so that often only parts of the mummies/skeletons were available for investigation. The available specimens were subjected to careful macroscopic examination, while isolated findings were radiologically analyzed. Using this approach, we identified at least 4 cases showing malignant tumors affecting the skeleton. In two cases, multiple mixed osteolytic-osteoblastic lesions suggested multiple metastases from carcinomas. Two further individuals presented with multiple osteolyses (vertebra, pelvis, skull) most suggestive of multiple myeloma. The observation of at least 4 cases of malignant tumors with osseous manifestation in a series of 325 adult individuals provides clear evidence that malignant tumors were not a rare event in the ancient Egyptian study population, particularly when the limitations of a study of tumors manifested only in osseous remnants are taken into consideration. A calculation of the age- and sex-adjusted tumor frequency in our material in comparison with a recent model for such a material by Waldron (1996) indicates that the rate of malignant tumors with bone affection in our series is higher than in an English population from 1901-1905, although lower than in a comparable present day population. This clearly indicates that important factors affecting malignant tumors were effective even in historic populations.


[The effect of serum starvation on the efficacy of the nuclear reprogramming of rabbit embryonic fibroblasts].
Successful transplantation of mammalian nuclei from differentiated cells has become possible after the application of original methods directed at the synchronization of cell cycles of the donor cell and recipient cytoplasm. We obtained a line of rabbit fetal fibroblasts which was used to study factors affecting the success of reprogramming. The nuclei of fetal fibroblasts (up to the 10th passage inclusive) proved to be capable of reprogramming and ensuring development of the cloned embryos until the preimplantation stages. The influence of synchronization of the cell cycles of the nucleus donor and recipient on the efficiency of reprogramming was studied. The rate of development of the cloned rabbit embryos to the morula-blastocyst stage reached 67% when the nuclei used were from stationary culture cells (G0-phase).


Early-Pregnancy Intermediate Hyperglycemia and Adverse Pregnancy Outcomes Among Women Without Gestational Diabetes.
Universal early-pregnancy screening for overt diabetes reveals intermediate hyperglycemia (fasting plasma glucose [FPG] [5.1-6.9 mM]). We evaluated the association between early-pregnancy intermediate hyperglycemia and adverse pregnancy outcomes among women without gestational diabetes. This retrospective cohort study was conducted at the Obstetrics and Gynecology Hospital, Shanghai, China, from 2013 to 2017. All singleton pregnancies with FPG less than or equal to 6.9 mM in early pregnancy and receiving a 75-g oral glucose tolerance test (OGTT) were included. Women with prepregnancy diabetes were excluded. Individuals with normal OGTT were analyzed. Pregnancy outcomes for FPG less than 5.1 mM and intermediate hyperglycemia were evaluated. The primary outcomes were large for gestational age (LGA) and primary cesarean delivery. Multivariate logistic regressions were conducted. Statistical significance was defined as P less than .05. In total, 24 479 deliveries were included, of which 23 450 (95.8%) had normal OGTTs later in pregnancy (NGT). There were 807 (3.4%) women who had an FPG of 5.1 to 6.9 mM in early pregnancy. Compared to the NGT group with an FPG of less than 5.1 mM in early pregnancy (N = 20692), the intermediate hyperglycemia NGT group (N = 693) had a higher age and body mass index (BMI), and significantly higher rates of LGA, primary cesarean delivery, preterm birth, preeclampsia, and neonatal distress. The rates of primary cesarean delivery (adjusted odds ratio [AOR] 1.24; 95% CI, 1.05-1.45), preterm birth (AOR 1.75; 95% CI, 1.29-2.36), and neonatal distress (AOR 3.29; 95% CI, 1.57-6.89) remained statistically significantly higher after adjustments for maternal age, BMI, and other potential confounding factors. Women with intermediate hyperglycemia in early pregnancy are at an increased risk for adverse maternal-fetal outcomes, even with normal future OGTTs.


Limitations of using the DATASUS database as a primary source of data in surgical research: a scoping review.
DATASUS is the Brazilian Public Unified Health System (SUS) department responsible for providing health data that are used as a primary source of data in several studies on surgery and surgical specialties although its main limitations have not been previously reviewed. The objective of this work is to synthesize information from studies on surgery that used DATASUS systems as a data source and to identify the main gaps in this platform. a scoping review was conducted according to the PRISMA-ScR method to identify papers on surgery, and other surgical specialties, that used the DATASUS platform as a primary data source. No restrictions were imposed regarding the type of study or year of publication. Grounded Theory was used to analyze the content of the articles. 248 works were initially analyzed and 47 were included in the final analysis of this study. The original articles included were published between 2009 and 2022 and the majority (12.76%, n=6) were published in the Journal of the Brazilian College of Surgeons. Retrospective studies (40.43%, n=19) were the most common type of study found. Content analysis of the articles identified four predominant domains in the scientific literature about the limitations of using DATASUS in surgical research: lack of data, reliability, precision and data integration. the information systems available in DATASUS are the largest source of information about the SUS, but the scientific literature on the quality of data available in these systems remains scarce and studies aimed at measuring this metric are necessary.


Technique of Hautmann ileal neobladder with chimney modification: interim results in 50 patients.
We report our 4-year experience with the chimney modification of the Hautmann ileal neobladder. This modification involves use of an 8 to 12 cm. tubularized isoperistaltic ileal chimney for the ureterointestinal anastomosis. Between April 1995 and March 1998, 50 men and women with invasive bladder cancer underwent radical cystectomy and creation of a Hautmann neobladder with chimney modification. Complications were assessed, divided as early and late, and subdivided as those related or unrelated to the neobladder. Continence was evaluated using a detailed patient questionnaire. There were no intraoperative deaths. Early complications in 11 of the 50 patients were neobladder related in 5 (10%) and unrelated to the neobladder in 6 (12%). The early reoperation rate was 6%. Late postoperative complications in 10 patients (20%) were neobladder related in 8 (16%) and unrelated to the neobladder in 2 (4%). After 1 year 93% and 86% of patients achieved good day and nighttime continence, respectively. In 2 patients (4%) clean intermittent catheterization is performed and 1 required placement of an artificial urinary sphincter. Ureterointestinal anastomotic strictures were detected in 6 of 100 ureteral units (6%), including 2 with failed initial endoscopic management. Open surgical revision of the ureterointestinal anastomotic site was easier due to the anterior position of the ureters, and identification and mobilization of the isoperistaltic limb. Our experience with the chimney modification of the Hautmann neobladder compares favorably to other forms of orthotopic urinary diversion in regard to ureteral stenosis, early and late postoperative complications, urinary continence and simplification of the ureterointestinal anastomosis.


Ureterolithotripsy in the management of calcified double-J catheter.
The authors present their experience in the management of a rare complication observed during long-term use of a ureteral catheter. The calcification along the entire surface of the catheter followed by incrustation can make its removal by means of cystoscopy difficult, thus becoming a great challenge to the urologist. The authors describe their experience in the management of this complication through the analysis of two illustrative cases. After failing to remove the catheter using extracorporeal lithotripsy, ureterolithotripsy was performed and the calcified ureteral catheter was successfully removed in both cases. Ureterolithotripsy proved to be an effective and safe method in the management of calcified double-J catheter.


The use of ultralow doses of antibodies to C-terminal fragment of angiotensin II AT1 receptor (kardos) in the therapy of arterial hypertension.
Kardos monotherapy allows attaining the target levels of systolic and diastolic blood pressure in patients with high-risk and very-high-risk hypertension. We demonstrated excellent tolerability of the preparation in combination with reliable blood pressure decrease over 24 h, during day and night hours.


[Green hair caused by frequent swimming pool use].
Three patients presented with an acquired green discoloration of their scalp hair. History revealed that all of them swam regularly in private swimming pools. Examination of the hair by atomic emission spectroscopy showed that the green discoloration was caused by an excessively high copper content of the hair. This exogeneous discoloration is characteristically related to the uptake of copper from private swimming pools.


Transport of Metabolites across Isolated Envelope Membranes of Spinach Chloroplasts.
Isolated envelope membranes of spinach chloroplasts (Spinacia oleracea L. var. Viroflay) exhibited selective permeability. Metabolites such as 3-phosphoglycerate, bicarbonate, glyoxylate, and acetate were transported rapidly; 6-phosphogluconate, glycolate, glycine, l-malate, and succinate were intermediate; whereas glucose 6-phosphate, fructose 1,6-diphosphate, and sucrose were hardly transported. Transport rates, metabolite accumulations within the membrane vesicles, and the internal water volume of isolated and in situ envelope membranes were compared and found to show similar trends.


[Role of P-JNK and P-c-Jun of JNK transduction pathway on the nasal mucosa remodeling in allergic rhinitis rats].
To study the role of P-JNK and P-c-Jun of JNK (c-Jun N-terminal kinase) on nasal mucosa remodeling in allergic rhinitis rats. Sixty male Wistar rats (weighing about 200-250 g) were randomly divided into AR group (A group) and B group(control group). The rats in A group were sensitized for inducing AR by intraperitoneal injection of ovalbumin and Al(OH)₃. Rats in group A were randomized into A4, A8 and A12 group (each had 10 rats). Ovalbumin was dropped in each nasal cavity of every rat for 4,8,12 weeks, respectively. Rats in group B were sensitized by saline instead of OVA, and were also divided into B4, B8 and B12 group. Each group had 10 rats. Pathological changes of nasal mucosa in each period were observed by hematoxylin and eosin stain dyeing. The phosphorylation of JNK and c-Jun were tested by immunohistochemistry. In A8 group, mucosal congestion and edema thickening with inflammatory cells infiltration of eosinophils were observed in the eighth week, and the inflammatory changes were significantly increased as time went on. The mean absorbance values of P-JNK and P-c-Jun in A group were significantly higher than those in the corresponding B group (all P < 0.01). Moreover, the mean absorbance values of A12 group were significantly higher than A4 group and A8 group (all P < 0.01 ). The expression of P-JNK and P-c-Jun in the process of nasal mucosa remodeling in allergic rhinitis rats were increased, which suggested that P-JNK and P-c-Jun played important roles in nasal mucosa remodeling of the allergic rhinitis rats.


A Retrospective Overview of Zika Virus Evolution in the Midwest of Brazil.
Since the introduction of the Zika virus (ZIKV) into Brazil in 2015, its transmission dynamics have been intensively studied in many parts of the country, although much is still unknown about its circulation in the midwestern states. Here, using nanopore technology, we obtained 23 novel partial and near-complete ZIKV genomes from the state of Goiás, located in the Midwest of Brazil. Genomic, phylogenetic, and epidemiological approaches were used to retrospectively explore the spatiotemporal evolution of the ZIKV-Asian genotype in this region. As a likely consequence of a gradual accumulation of herd immunity, epidemiological data revealed a decline in the number of reported cases over 2018 to 2021. Phylogenetic reconstructions revealed that multiple independent introductions of the Asian lineage have occurred in Goiás over time and revealed a complex transmission dynamic between epidemic seasons. Together, our results highlight the utility of genomic, epidemiological, and evolutionary methods to understand mosquito-borne epidemics. IMPORTANCE Despite the considerable morbidity and mortality of arboviral infections in Brazil, such as Zika, chikungunya, dengue fever, and yellow fever, our understanding of these outbreaks is hampered by the limited availability of genomic data to track and control the epidemic. In this study, we provide a retrospective reconstruction of the Zika virus transmission dynamics in the state of Goiás by analyzing genomic data from areas in Midwest Brazil not covered by other previous studies. Our study provides an understanding of how ZIKV initiates transmission in this region and reveals a complex transmission dynamic between epidemic seasons. Together, our results highlight the utility of genomic, epidemiological, and evolutionary methods to understand mosquito-borne epidemics, revealing how this toolkit can be used to help policymakers prioritize areas to be targeted, especially in the context of finite public health resources.


Tubulointerstitial macrophage infiltration in a patient with hypokalemic nephropathy and primary Sjögren's syndrome.
We report a case of hypokalemic nephropathy associated with primary Sjögren's syndrome (SS). The patient presented with profound and persistent hypokalemia secondary to distal renal tubular acidosis (RTA). A renal biopsy exhibited tubular degeneration, marked interstitial fibrosis and intense macrophage infiltration. Hypokalemia has been reported to induce macrophage infiltration in experimental animal models but not in humans. This is the first report of intense tubulointerstitial macrophage infiltration in a patient with hypokalemic nephropathy associated with SS.


An improved snare system for the nonsurgical retrieval of intravascular foreign bodies.
We have modified a snare technique originally described for transcolonoscopic removal of colonic polyps for the retrieval of intravascular foreign bodies. The key difference from other snare techniques is the employment of a wire snare with a crimp in its midportion that enables the formation of a loop in a plane perpendicular to the axial direction of the guiding catheter. With this technique we have quickly removed a variety of objects from the right heart and pulmonary arteries in four patients.


Transportable lunar-ranging system.
Ruby lasers in large, fixed installations are being used by groups in France, Japan, the Soviet Union, and the U.S. for measuring distances to retroreflectors on the moon. This paper describes a transportable transmitting unit that can be installed at any astronomical observatory where a large telescope is available to detect the received signal. The transmitter consists of a high radiance, frequency-doubled, neodymium-glass laser and a coudé optical system of moderate size. It has been installed, but is not yet in operation, at the Agassiz Observatory, Harvard, Massachusetts.


[Expression of vascular endothelial growth factor C in human breast cancer cell lines MCF-7 and MCF-7/Adr].
Vascular endothelial growth factor C (VEGF-C) discovered recently is a new member of vascular endothelial growth factor (VEGF) family. Studies have shown that VEGF-C is one specific regulating factor for lymphatic endothelial cells, and it expresses in a variety of human tumors. In order to explore the role of VEGF-C in the process of human tumor formation and development, the authors designed this study to examine the expressions of VEGF-C mRNA and protein in human breast cancer cell line MCF-7 and drug resistant cell line MCF-7/Adr. According to the VEGF-C gene sequence, A specific oligonucleotide probe labeled with digoxin was designed and constructed; Then in situ hybridization was applied to detect the expression of VEGF-C mRNA in the cultured human breast cancer cell lines MCF-7 and MCF-7/Adr respectively; Finally immunohistochemistry was performed to examine the expression of VEGF-C protein in both cell lines. Blue positive staining granules for VEGF-C mRNA were observed in the cytoplasm of these cells by in situ hybridization where brown positive staining granules for VEGF-C protein were also observed by immunohistochemistry. While there were no positive staining granules in neither control cells. Human breast cancer cell lines MCF-7 and MCF-7/Adr can transcribe VEGF-C mRNA and translate corresponding protein in their cytoplasm.


The cross-priming APC requires a Rel-dependent signal to induce CTL.
Induction of OVA-specific CTL by cross-priming requires help from CD4 T cells, which use CD154 to signal CD40 on the APC. To further dissect the molecular pathways involved in cross-priming, we examined the role of Rel, an NF-kappaB family member. c-rel(-/-) mice failed to generate OVA-specific CTL by cross-priming, but could induce CTL to HSV-1. Using chimeric mice, Rel expression was shown to be required by the APC, but not by the T cells. Notably, the deficiency in Rel could be overcome by triggering CD40, implying that the APC required Rel before receipt of the CD40 signal. These data suggest that the cross-priming APC must receive two signals before it can stimulate CTL. The first signal is Rel dependent and is required before activation of CD4 helper T cells, which then deliver the second signal using CD154 to trigger CD40.


Identifying a Single Optimal Integrated Cervical Cancer Prevention Policy in Norway: A Cost-Effectiveness Analysis.
Interventions targeting the same disease but at different points along the disease continuum (e.g., screening and vaccination to prevent cervical cancer [CC]) are often evaluated in isolation, which can affect cost-effectiveness profiles and policy conclusions. We evaluated nonavalent human papillomavirus (HPV) vaccine (9vHPV) compared with bivalent HPV vaccine (2vHPV) alongside deintensified screening intervals for a vaccinated birth cohort to inform a single optimal integrated CC prevention policy. Using a multimodeling approach, we evaluated the health and economic impacts of alternative CC screening strategies for a Norwegian birth cohort eligible for HPV vaccination in 2021 assuming they received 1) 2vHPV or 2) 9vHPV. We conducted 1) a restricted analysis that evaluated the optimal HPV vaccine under current screening guidelines; and 2) a comprehensive analysis including alternative screening and vaccination strategy combinations. We calculated incremental cost-effectiveness ratios (ICERs) and evaluated them according to different cost-effectiveness thresholds. Assuming a cost-effectiveness threshold of $40,000 per quality-adjusted life year (QALY) gained, we found that, while holding screening intensity fixed, switching the routine vaccination program in Norway from 2vHPV to 9vHPV would not be considered cost-effective (ICER of $132,700 per QALY gained). However, when allowing for varying intensities of CC screening, we found that switching to 9vHPV would be cost-effective compared with 2vHPV under an alternative threshold of $55,000 per QALY gained, if coupled with reductions in the number of lifetime screens. Our analysis highlights the importance of evaluating the full potential policy landscape for country-level decision makers considering policy adoption, including nonindependent primary and secondary prevention efforts, to draw appropriate conclusions and avoid sub-optimal outcomes. Without evaluating the full potential policy landscape, including primary and secondary prevention efforts, country-level decision makers may not be able to draw appropriate policy conclusions, resulting in suboptimal outcomes.An applied example from cervical cancer prevention in Norway compared a restricted analysis of current screening guidelines to a comprehensive analysis including alternative screening and vaccination strategy combinations.We found that a switch from bivalent to nonavalent human papillomavirus vaccine would be considered cost-effective in Norway if coupled with reductions in the number of lifetime screens compared with the current screening strategy.A comprehensive analysis that considers how different types of interventions along the disease continuum affect each other will be critical for decision makers interpreting cost-effectiveness analysis results.


Potential Risk Factors for Isolated Hypothyroxinemia in Women of Childbearing Age-Results from Retrospective Analysis.
Isolated hypothyroxinemia (IH) unfavorably affects reproduction. This study aimed to evaluate retrospectively if any routinely measured clinical/laboratory parameters are associated with IH among women of childbearing age hospitalized in the endocrine department. A group of 466 female non-pregnant inpatients (age range 13-57 years) was considered. IH (decreased free thyroxine (FT4) with normal TSH) was found in 8/466 patients (1.72%). Vitamin D deficiency (<30 ng/mL) was found in all patients with IH, whereas severe Vitamin D deficiency (<20 ng/mL) was found in 5/6. Vitamin D concentration was lower in IH females. FT4 concentration was lower in patients with severe vitamin D deficiency and correlated positively with vitamin D concentration. Insulin resistance index (IRI) was increased (>1.25) in 5/6 patients with IH. IRI was higher in IH patients and it was the only independent linear factor for IH in the univariate regression. FT4 concentration was lower in patients with increased IRI and correlated negatively with IRI. FT4 concentration correlated negatively with body mass index (BMI) and LDL cholesterol or triglycerides, and positively with HDL cholesterol or HDLC/cholesterol ratio. Vitamin D deficiency, insulin resistance and increased BMI (as potential causative factors), and abnormal lipid profile (as a possible consequence), are associated with IH in women of childbearing age. Eliminating risk factors for hypothyroxinemia may improve reproductive health.


Genetic differentiation of some Glossina morsitans morsitans populations.
To study the population structure of Glossina morsitans morsitans Westwood (Diptera: Glossinidae), polymerase chain reaction (PCR) and singlestrand conformational polymorphism (SSCP) methods were used to estimate mitochondrial DNA diversity at four loci in six natural populations from Zambia, Zimbabwe and Mozambique, and in two laboratory cultures. The Zambian and Zimbabwean samples were from a single fly belt. Four alleles were recorded at 12S and 16S1, and five alleles at 16S2 and COI. Nucleotide sequencing confirmed their singularities. Chi-square contingency tests showed that allele frequencies differed significantly among populations. Mean allele diversities in populations averaged over loci varied from 0.14 to 0.61. Little loss in haplotype diversity was detected in the laboratory cultures thereby indicating little inbreeding. Wright's fixation index F(ST) in the natural populations was 0.088+/-0.016, the correlation of haplotypes within populations relative to correlations in the total. A function of its inverse allows an estimate of the mean equivalent number of females exchanged per population per generation, 5.2. No correlation was detected between pairwise genetic distance measures and geographical distances. Drift explains the high degree of differentiation.


Temperature increase during orthodontic bonding with different curing units using an infrared camera.
To evaluate the effects of different curing units and light-tip tooth surface distances on the temperature increase generated during orthodontic bonding, using an infrared camera (IR) and artificial neural networks (ANN). Fifty-two freshly extracted human premolar teeth were used. Metallic orthodontic brackets were bonded to the buccal surfaces of the teeth and thermal records were taken using an IR camera and ANN. Brackets were cured with a light-emitting diode (LED) and high intensity halogen (HQTH). Teeth were divided into four groups according to the curing units (LED and HQTH) and curing distances (from tooth surface and 10 mm away from tooth surface). The results were analyzed with analysis of variance (ANOVA) and the Tukey HSD test. The ANOVA and Tukey HSD tests revealed that temperature changes were influenced by the type of light source and exposure times. All groups revealed significant differences between each other (p < 0.001). The highest surface temperature increase was gained from curing with a LED unit from the tooth surface (11.35°C ± 0.91°C). The lowest surface temperature increase was gained from curing with a HQTH unit 10 mm away from the tooth surface (2.57°C ± 0.6°C). The LED unit induced significantly higher temperature changes than did the HQTH. The temperature increase during orthodontic bonding was increased with long exposure time. A shorter light-tip tooth surface distance leads to greater increases in temperature.


HIV-induced type I interferon and tryptophan catabolism drive T cell dysfunction despite phenotypic activation.
Infection by the human immunodeficiency virus (HIV) is characterized by functional impairment and chronic activation of T lymphocytes, the causes of which are largely unexplained. We cultured peripheral blood mononuclear cells (PBMC) from HIV-uninfected donors in the presence or absence of HIV. HIV exposure increased expression of the activation markers CD69 and CD38 on CD4 and CD8 T cells. IFN-alpha/beta, produced by HIV-activated plasmacytoid dendritic cells (pDC), was necessary and sufficient for CD69 and CD38 upregulation, as the HIV-induced effect was inhibited by blockade of IFN-alpha/beta receptor and mimicked by recombinant IFN-alpha/beta. T cells from HIV-exposed PBMC showed reduced proliferation after T cell receptor stimulation, partially prevented by 1-methyl tryptophan, a competitive inhibitor of the immunesuppressive enzyme indoleamine (2,3)-dioxygenase (IDO), expressed by HIV-activated pDC. HIV-induced IDO inhibited CD4 T cell proliferation by cell cycle arrest in G1/S, and prevented CD8 T cell from entering the cell cycle by downmodulating the costimulatory receptor CD28. Finally, the expression of CHOP, a marker of the stress response activated by IDO, was upregulated by HIV in T cells in vitro and is increased in T cells from HIV-infected patients. Our data provide an in vitro model for HIV-induced T cell dysregulation and support the hypothesis that activation of pDC concomitantly contribute to phenotypic T cell activation and inhibition of T cell proliferative capacity during HIV infection.


Machine-Learning-Based Approach to Differential Diagnosis in Tuberculous and Viral Meningitis.
Tuberculous meningitis (TBM) is the most severe form of tuberculosis, but differentiating between the diagnosis of TBM and viral meningitis (VM) is difficult. Thus, we have developed machine-learning modules for differentiating TBM from VM. For the training data, confirmed or probable TBM and confirmed VM cases were retrospectively collected from five teaching hospitals in Korea between January 2000 - July 2018. Various machine-learning algorithms were used for training. The machine-learning algorithms were tested by the leave-one-out cross-validation. Four residents and two infectious disease specialists were tested using the summarized medical information. The training study comprised data from 60 patients with confirmed or probable TBM and 143 patients with confirmed VM. Older age, longer symptom duration before the visit, lower serum sodium, lower cerebrospinal fluid (CSF) glucose, higher CSF protein, and CSF adenosine deaminase were found in the TBM patients. Among the various machine-learning algorithms, the area under the curve (AUC) of the receiver operating characteristics of artificial neural network (ANN) with ImperativeImputer for matrix completion (0.85; 95% confidence interval 0.79 - 0.89) was found to be the highest. The AUC of the ANN model was statistically higher than those of all the residents (range 0.67 - 0.72, P <0.001) and an infectious disease specialist (AUC 0.76; P = 0.03). The machine-learning techniques may play a role in differentiating between TBM and VM. Specifically, the ANN model seems to have better diagnostic performance than the non-expert clinician.


Role of carbon monoxide dehydrogenase in acetate synthesis by the acetogenic bacterium, Acetobacterium woodii.
Carbon monoxide dehydrogenase (CODH) plays a key role in acetate synthesis by the acetogenic bacterium, Clostridium thermoaceticum. Acetobacterium woodii, like C. thermoaceticum contains high levels of CODH. In this work we show that crude extracts of A. woodii synthesize acetate from methyl tetrahydrofolate or methyl iodide, carbon monoxide and coenzyme A (CoA). The purified CODH from A. woodii catalyzes an exchange reaction between CO and the carbonyl group of acetyl-CoA even faster than the C. thermoaceticum enzyme, indicating the CODH of A. woodii, like that of C. thermoaceticum is an acetyl-CoA synthetase. Fluorescence and EPR studies further support this postulate by demonstrating that CODH binds CoA near the CO binding site involving a tryptophan residue. The UV absorption spectra and the amino acid compositions of A. woodii and C. thermoaceticum CODHs are very similar. Evidence is presented using purified enzymes from A. woodii that the synthesis of acetyl-CoA occurs by a pathway similar to that utilized by C. thermoaceticum.


A Constitutive Relationship for Gravelly Soil Considering Fine Particle Suffusion.
Suffusion erosion may occur in sandy gravel dam foundations that use suspended cutoff walls. This erosion causes a loss of fine particles, degrades the soil strength and deformation moduli, and adversely impacts the cutoff walls of the dam foundation, as well as the overlying dam body. A comprehensive evaluation of these effects requires models that quantitatively describe the effects of fine particle losses on the stress-strain relationships of sandy gravels. In this work, we propose an experimental scheme for studying these types of models, and then perform triaxial and confined compression tests to determine the effects of particle losses on the stress-strain relationships. Considering the Duncan-Chang E-B model, quantitative expressions describing the relationship between the parameters of the model and the particle losses were derived. The results show that particle losses did not alter the qualitative stress-strain characteristics of the soils; however, the soil strength and deformation moduli were degraded. By establishing the relationship between the parameters of the model and the losses, the same model can then be used to describe the relationship between sandy gravels and erosion levels that vary in both time and space.


[Create or copy... Which is the difference?].
Creating and copying are two different processes; we must not confuse creativity with plagiarism. However, this distinction is problematic, because there is no possibility of creating from scratch, this implies that any creative act necessarily arises from accumulative experience, inevitably producing a continuity between old and new. Even so it is necessary to establish clearly the difference between creating and copying. It is not desirable that a matter of such importance remains in the nebula or that the relationship between creativity and ethics is kept unaware. There are many cases of plagiarism, but this cannot be a consolation. There is no gain when the existence of a plagiarism is ignored or concealed and less when it is unjustified.


Role of CD4 hinge region in GP120 utilization by immunoglobulin domain 1.
Immunoglobulin-like domain 1 of CD4 (D1-CD4) promotes HIV infection by binding the envelope glycoprotein (ENV) and exposing its coreceptor-binding site. To study CD4-ENV-coreceptor interactions, we characterized hybrid receptors having domains 1 and 2 of CD4 (D1D2-CD4) joined to the N-terminus of chemokine receptors CCR5, CXCR4, CXCR2, and DARC. Hybrid receptors showed conserved ENV-coreceptor specificity in cell-cell fusion assays. Although D1D2-CD4-CCR5 was sufficient to permit ENV-mediated fusion, D1-CD4-CCR5 and human D1/mouse D2-CD4-CCR5 lacked CD4 function and binding to a neutralizing antibody mapped to D1-CD4. Chimeric D1D2-CD4 joined to CCR5 revealed that the C-terminal 20 residues of human D2-CD4 are required for efficient ENV-mediated fusion. Mutagenesis of hybrid receptors showed the importance of residues forming D1-D2 CD4 interdomain contacts and hinge region proximal residues. Mutagenesis of WT human CD4 confirmed that residues forming D1-D2 interdomain contacts and hinge-region proximal residues contribute positively to CD4 activity in the full-length receptor.


Detection of Salmonella in chicken meat by insulated isothermal PCR.
Consumption of Salmonella-contaminated foods, such as poultry and fresh eggs, is known to be one of the main causes of salmonellosis. Conventional PCR methods, including real-time PCR for rapid detection of Salmonella, in general require skilled technicians and costly instruments. A recently developed novel convective PCR, insulated isothermal PCR (iiPCR), is carried out in polycarbonate capillary tubes. In this study, we designed TaqMan probes and PCR primers based on the yrfH gene encoding a heat shock protein for the iiPCR detection of Salmonella in chicken meat samples. The TaqMan probe was labeled with 6-carboxyfluorescein and 6-carboxytetramethylrhodamine at the 5' and 3' ends, respectively. The PCR amplicon was 133 bp. A typical run of this iiPCR assay was completed within 1 h. Specific PCR products were obtained for 148 strains representing 49 serotypes of Salmonella tested. Under the same conditions, false-positive results were not obtained for 98 non-Salmonella strains tested, including strains of Enterobacteriaceae closely related to Salmonella. For chicken meat samples, with a 5-h enrichment step Salmonella at as low as 10⁰ CFU/g of poultry meat could be detected. Because the amplification signals from the probes are detectable at 520 nm, identification of the PCR products by gel electrophoresis is not required. Compared with conventional PCR, the iiPCR system requires less expertise and provides an economical, reliable, and rapid tool for result interpretation. Detection results can be obtained within 8 h, including the enrichment and DNA extraction steps.


A free-choice high-fat high-sugar diet induces changes in arcuate neuropeptide expression that support hyperphagia.
The mechanisms for how saturated fat and sugar-based beverages contribute to human obesity are poorly understood. This paper describes a series of experiments developed to examine the response of hypothalamic neuropeptides to diets rich in sugar and fat, using three different diets: a high-fat high-sugar (HFHS) choice diet with access to chow, saturated fat and a 30% sugar solution; a high-fat (HF) choice diet with access to chow and saturated fat; or to a high-sugar (HS) choice diet with access to chow and a sugar solution. We first studied caloric intake, body weight gain, hormonal alterations and hypothalamic neuropeptide expression when male Wistar rats were subjected to an HFHS choice, an HF choice or an HS choice diet for 1 week. Next, we studied caloric intake and body weight gain when rats were subjected to the choice diets for 5 weeks. Finally, we measured neuropeptide expression in hepatic vagotomized rats subjected to an HFHS choice, an HF choice or an HS choice diet for 1 week. In rats on an HF choice diet, plasma leptin concentrations and proopiomelanocortin (POMC) mRNA increased and neuropeptide Y (NPY) mRNA decreased. Rats on an HFHS choice diet showed identical plasma leptin concentrations as rats on an HF choice diet. However, NPY mRNA increased and POMC mRNA decreased. An HS choice diet for 1 week did not alter hypothalamic neuropeptide expression or plasma leptin concentrations. As hormonal changes did not explain the differences in hypothalamic neuropeptide expression between rats on the choice diets, we addressed whether neuronal feedback signals mediated the hypothalamic neuropeptide response. The POMC mRNA response to different diets depended on an intact innervation of liver and upper intestinal tract. Our data suggest that the specific combination of saturated fat and a 30% sugar solution results in hyperphagia-induced obesity and alters hypothalamic neuropeptide expression, and that the response of the melanocortin system is mediated by the hepatic vagus.


The psychological and behavioural correlates of COVID-19 vaccine hesitancy and resistance in Ireland and the UK.
The successful control of the COVID-19 pandemic depends largely on the acceptance and uptake of a COVID-19 vaccine among the public. Thus, formative research aiming to understand and determine the causes of weak and/or positive vaccination intentions is vital in order to ensure the success of future and current vaccination programmes through the provision of effective, evidence-based health messaging. A cross-sectional survey was completed by a sample of Irish (N = 500) and UK (N = 579) citizens using the online platform 'Qualtrics'. Participants completed a questionnaire battery comprised of health, attitudes/beliefs, influences, and behavioural intention measures. Demographic information was also assessed. Results highlighted similar rates of vaccine intention among both samples; where a total of 76.8% Irish respondents, and 73.7% of UK respondents indicated that they intended to be immunized if the government advised them to take the COVID-19 vaccine. Overall, 23.2% of Irish respondents reported being vaccine hesitant or vaccine resistant, while a rate of 26.3% of UK respondents reported vaccine hesitancy or resistance. Univariate analysis highlighted that both gender and age played a significant role in vaccine intention, with women under age 30 reporting higher rate of vaccine hesitancy. Multivariate analysis revealed that significant correlates of vaccine acceptance included peer influence, GP influence, civic responsibility, perceived benefit, and positive vaccination attitudes. Those who reported vaccine resistance and hesitancy were more likely to have less positive vaccination attitudes and perceive higher vaccination risk. The current sociodemographic and psychological profiles of vaccine resistant and hesitant individuals provide a useful resource for informing health practitioners in the UK and Ireland with the means of enhancing pro-vaccine attitudes and promoting vaccination uptake. The current research shows indications of associations between distrust in the vaccine itself and vaccine hesitancy and resistance. Thus, to effectively design and deliver public health messages that ensures the success of vaccination uptake, it is likely that governments and public health officials will need to take actions to garner trust in the safety of the vaccine itself. Additionally, campaigns to decrease hesitancy and resistance in the COVID-19 vaccine may benefit in targeting altruism to increase willingness to get vaccinated against COVID-19.


Self-Induced Euglycemic Diabetic Ketoacidosis: When to Stop the Drip.
Diabetic ketoacidosis (DKA) is a well-known, serious complication that many patients with type 1 and 2 diabetes face due to either a relative or absolute insulin deficiency. Sodium-glucose cotransporter 2 (SGLT-2) inhibitors have gained increased popularity due to their diabetic, cardiovascular, and renal benefits. An associated complication of SGLT2 inhibitors is euglycemic DKA. A 56-year-old male with a history of type 2 diabetes mellitus and peripheral neuropathy presented with right foot pain secondary to a diabetic foot ulcer. The ulcer was present for one year, but the patient noticed increased pain and purulent discharge over the three days prior to presentation. While being treated inpatient for the foot ulcers, the patient repeatedly refused to receive standard hospital diabetes management of insulin injections. He instead insisted to take his home medications against medical advice, which were metformin and Glyxambi® (empagliflozin/linagliptin, Boehringer Ingelheim, Ingelheim am Rhein, Germany). His diabetic foot ulcer was medically managed with IV antibiotics. On day 4 of admission, his anion gap increased to 23 mEq/L, and serum bicarbonate (CO2) decreased to 8 mEq/L, raising concerns of diabetic ketoacidosis. His glucose was 141 mg/dL, his beta-hydroxybutyrate was high at 5.5 mmol/L. An arterial blood gas (ABG) test demonstrated anion gap metabolic acidosis with secondary respiratory alkalosis. A urinalysis demonstrated glucose 1000 mg/dL and ketones of 150 mg/dL. The patient was diagnosed with euglycemic DKA. Due to the patient having normal glucose levels, an insulin drip and a 5% dextrose with 0.45% normal saline drip were started. Basic metabolic profiles were ordered every four hours, with glucose checks every hour. Once the anion gap was closed and his urinary ketones disappeared, the patient transitioned to subcutaneous insulin. He was able to be discharged home with basal subcutaneous insulin and metformin with instructions to avoid SGLT2 inhibitors in the future. Unfortunately, there are currently no guidelines from endocrinology or internal medicine societies regarding the management of euglycemic DKA. As the typical DKA diagnostic criteria of elevated blood glucose level are not present, it is easy to overlook euglycemic DKA. As these SGLT2 inhibitors become more prevalent, careful monitoring of all potential side effects as well as the contraindications are prudent to successful management of complex disease states.


Strategic planning in competitive environments.
Private voluntary social welfare organizations (PVSOs) are operating under increasingly competitive conditions. "Strategic Planning" (SP) is a management tool designed to meet those conditions. In an exploratory study of 154 PVSOs the authors examined management's reasons for adopting strategic planning, the attention given to external analysis, and the degree of change and satisfaction achieved. PVSOs use SP because they are required/urged to do so by external agencies. External analysis received the least attention. One-half of the organizations achieved major change which was associated with high levels of satisfaction. Widespread stakeholder participation and the use of consensus strategies detracted from both major change and high satisfaction outcomes.


Immunohistochemical Evaluation of BAP1 Expression in Breast Cancer with Known BRCA1 and BRCA2 Mutations and Comparison with Histopathological Features.
Introduction. BRCA-mutated breast cancers have specific pathological characteristics. BAP1 is a tumor suppressor gene that is important in many cancers with different pathways. The relationship between BRCA1 mutation and BAP1 immunohistochemical staining is still unclear. Our aim is to determine whether BAP1 immunohistochemical expression indicates BRCA mutation status in breast carcinomas with specific pathological characteristics. In addition, we aim to determine the histopathological characteristics of tumors according to BRCA mutations. Methods. Histomorphology, molecular subtypes and BAP1 immunohistochemical expression patterns of the BRCA1/BRCA2 mutated and non-mutated tumors were evaluated. The BAP1 immunohistochemical stain was applied to nine tumor tissues with the BRCA1 mutation, six tumor tissues with the BRCA2 mutation, and 16 tumor tissues without any BRCA mutation. Pearson's chi square test and the Fisher Freeman Halton test were used to analyze the associations between the datas. The statistical significance was considered as P value of <.05. Results. Immunohistochemical BAP1 loss was not detected in any mutated or non-mutated tumor group. BRCA1 mutated tumors had the statistically highest histopathological grade (P = .04) and BRCA1/2 mutated tumors had significant immunohistochemical triple negative expression pattern (P = .01). Conclusions. Intrinsic and histopathological characteristics may vary between BRCA1 mutated and non-BRCA1 mutated tumors. Also, BAP1 loss was not detected in BRCA mutated breast tumors because of several effects of BAP1 that are non-related with BRCA in the cell cycle.


Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., three new species of ballistoconidium-forming yeasts in the Agaricostilbum lineage isolated from plants in Taiwan.
Three strains of xylose-lacking and ubiquinone-10-having ballistoconidium-forming yeasts isolated from plant leaves collected in Taiwan were found to represent respective new species. In phylogenetic trees constructed based on the nucleotide sequences of 18S rDNA and D1/D2 domain of 26S rDNA, they were located in the Agaricostilbum lineage (Agaricostilbum/Bensingtonia cluster). Since the taxonomic properties of these species coincide with those of the genus Sporobolomyces, they are described as Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., respectively.


The Arabidopsis transcription factor WRKY27 influences wilt disease symptom development caused by Ralstonia solanacearum.
WRKY transcription factors play a key role in modulating the plant defense transcriptome. Here we show that the Arabidopsis mutant wrky27-1, which lacks a functional WRKY27 transcription factor, showed delayed symptom development in response to the bacterial wilt pathogen Ralstonia solanacearum. Additionally, wrky27-1 plants did not express PR marker genes upon infection, as also observed in resistant Nd-1 plants. Spatial expression of WRKY27 correlated well with the route of bacterial infection and propagation in planta. Complementation experiments restored both the early wilting phenotype of wild-type Col-1 plants and activation of PR genes, not only when the WRKY27 cDNA is expressed under the control of the native promoter, but also when the SUC2 promoter was used, suggesting that WRKY27 exerts its function in phloem companion cells. Expression studies identified genes involved in nitrogen metabolism and nitric oxide (NO) generation as potential targets of negative regulation by WRKY27. Our results show that WRKY27 negatively influences symptom development of a vascular pathogen, possibly by affecting signaling or trafficking between the phloem and the xylem.


Mask versus nasal tube for stabilization of preterm infants at birth: a randomized controlled trial.
Positive-pressure ventilation (PPV) using a manual ventilation device and a face mask is recommended for compromised newborn infants in the delivery room (DR). Mask ventilation is associated with airway obstruction and leak. A nasal tube is an alternative interface, but its safety and efficacy have not been tested in extremely preterm infants. An unblinded randomized controlled trial was conducted in Australia, and the Netherlands. Infants were stratified by gestational age (24-25/26-29 weeks) and center. Immediately before birth infants were randomly assigned to receive PPV and/or continuous positive airway pressure with either a nasal tube or a size 00 soft, round silicone mask. Resuscitation protocols were standardized; respiratory support was provided using a T-piece device commencing in room air. Criteria for intubation included need for cardiac compressions, apnea, continuous positive airway pressure >7 cm H2O, and fraction of inspired oxygen >0.4. Primary outcome was endotracheal intubation in the first 24 hours from birth. Three hundred sixty-three infants were randomly assigned; the study terminated early on the grounds of futility. Baseline variables were similar between groups. Intubation rates in the first 24 hours were 54% and 55% in the nasal tube and face mask groups, respectively (odds ratio: 0.97; 95% confidence interval: 0.63-1.50). There were no important differences in any of the secondary outcomes within the whole cohort or between the 2 gestational age subgroups. In infants at <30 weeks' gestation receiving PPV in the DR, there were no differences in short-term outcomes using the nasal tube compared with the face mask.


[The classification of atypical forms of neurofibromatosis].
Neurofibromatosis (NF) represents not an entity but a group of several forms which differ as to symptoms, prognosis and inheritance. In 1982 Riccardi suggested a classification into eight categories. Two of these--Von Recklinghausen NF-1 and bilateral acoustic NF-2- were defined in 1987 by a National Institutes of Health Consensus Development Conference. We describe 13 patients whose symptoms do not fit the diagnostic criteria for NF-1 and NF-2. 6 subjects can be classified as segmental NF-V. One of these patients was remarkable in having iris hamartomata, while his daughter was affected with NF-1. Another (female) patient in this group had areolar freckling, a finding not yet reported in NF patients. 4 cases belong to the late-onset NF-VII category, 3 of whom developed a neurofibrosarcoma. Two subjects are fist-degree relatives (mother and son). We are not aware of familial occurrence of NF-VII. Two further subjects were assigned to the variant form of NF, NF-IV. One patient had symptoms similar to NF-2 but the minimal diagnostic criteria were not fulfilled. Nevertheless, we consider his classification as NF-2 in view of a pattern of intracranial calcification repeatedly and exclusively found in NF-2. In general, assignment to a category of NF is important for appropriate patient management and genetic counselling. Classification of individual patients may be arbitrary at the time.


Response surface methodological optimization of L-asparaginase production from the medicinal plant endophyte Acinetobacter baumannii ZAS1.
This study targets the enhanced production of L-asparaginase, an antitumor enzyme by Acinetobacter baumannii ZAS1. This organism is an endophyte isolated from the medicinal plant Annona muricata. Plackett-Burman design (PBD) and central composite design (CCD) were used for statistical optimization of media components. The organism exhibited 18.85 ± 0.2 U/mL enzyme activities in unoptimized media. Eight variables: L-asparagine, peptone, glucose, lactose, yeast extract, NaCl, MgSO4, and Na2HPO4 were screened by PBD. Among them, only four factors-L-asparagine, peptone, glucose, and Na2HPO4-were found to affect enzyme production significantly (p < 0.05). Furthermore, the best possible concentrations and interactive effects of the components that enhance this enzyme's output were chosen by using CCD on these selected variables. The results revealed that an optimized medium produces a higher concentration of enzymes than the unoptimized medium. After optimizing media components, the maximum L-asparaginase activity was 45.59 ± 0.36 U/mL, around the anticipated value of 45.04 ± 0.42 U/mL. After optimization of process parameters, it showed a 2.41-fold increase in the production of L-asparaginase by the endophyte Acinetobacter baumannii ZAS1. The findings of this study indicated that an endophyte, Acinetobacter baumannii ZAS1 that produces L-asparaginase could be used to increase enzyme output. However, using the statistical methods Plackett-Burman design and central composite design of response surface methodology is a handy tool for optimizing media components for increased L-asparaginase synthesis.


MdWRKY126 modulates malate accumulation in apple fruit by regulating cytosolic malate dehydrogenase (MdMDH5).
The content of organic acids greatly influences the taste and storage life of fleshy fruit. Our current understanding of the molecular mechanism of organic acid accumulation in apple (Malus domestica) fruit focuses on the aluminum-activated malate transporter 9/Ma1 gene. In this study, we identified a candidate gene, MdWRKY126, for controlling fruit acidity independent of Ma1 using homozygous recessive mutants of Ma1, namely Belle de Boskoop "BSKP" and Aifeng "AF." Analyses of transgenic apple calli and flesh and tomato (Solanum lycopersicum) fruit demonstrated that MdWRKY126 was substantially associated with malate content. MdWRKY126 was directly bound to the promoter of the cytoplasmic NAD-dependent malate dehydrogenase MdMDH5 and promoted its expression, thereby enhancing the malate content of apple fruit. In MdWRKY126 overexpressing calli, the mRNA levels of malate-associated transporters and proton pump genes also significantly increased, which contributed to the transport of malate accumulated in the cytoplasm to the vacuole. These findings demonstrated that MdWRKY126 regulates malate anabolism in the cytoplasm and coordinates the transport between cytoplasm and vacuole to regulate malate accumulation. Our study provides useful information to improve our understanding of the complex mechanism regulating apple fruit acidity.


Lymph node metastases in papillary thyroid cancer detected by quantitative real-time polymerase chain reaction for thyroglobulin and cytokeratine-19.
Papillary thyroid cancer (PTC) metastases in the lymph nodes (LNs) were detected by real-time polymerase chain reaction (PCR) for TG and cytokeratin 19 (CK19), and the obtained results were compared with histopathology. 107 LNs from 34 PTC patients were divided into four blocks by a special cutting device - 2 for histopathology, while the other 2 were tested by quantitative real-time PCR. Metastases were detected in 20 nodes from 10 (29.4%) patients. TG and CK19 expression levels differed vastly between nodes with and without metastatic cells. ddCt of TG in the genetic material extracted from N0 nodes was 9.97 ±4.20, while in nodes with metastases ddCt was 0.91±4.20 (p < 0.0001). Cytokeratin 19 showed similar results with expression level (ddCt) in N0 nodes of 10.96 ±2.58 vs. 7.73 ±3.63 in nodes with metastases (p < 0.0001). Evaluation of the utility of both parameters showed efficient differentiation of node involvement in the case of TG, with area under the ROC curve (AUC) equal to 0.91 (95% CI: 0.85-0.96). Cytokeratin 19 also allowed for a degree of differentiation but its diagnostic efficacy was lower (AUC 0.76, 95% CI: 0.64-0.88). The combined TG and CK19 quantitative real-time PCR could be used to select a previously missed group of patients with nodal involvement undetectable by standard histopathology.