Wine aroma evolution throughout alcoholic fermentation sequentially inoculated with non- Saccharomyces/Saccharomyces yeasts.
Nine non-Saccharomyces yeasts belonging to 6 species (Torulaspora delbrueckii, Metschnikowia pulcherrima, Lachancea thermotolerans, Zygosaccharomyces bailii, Williopsis pratensis and Candida zeylanoides) and two mixed inoculum of T. delbrueckii and L. thermotolerans were screened for aroma formation and fermentative behaviour in sequential inoculations with Saccharomyces cerevisiae. The main differences in aromatic composition within wines were detected in the first stages of vinification between S. cerevisiae and non-Saccharomyces species, but not within the latter species. Most of the wines made with non-Saccharomyces in sequential fermentation with S. cerevisiae tended to produce higher ethanal and glycerol and lower volatile acidity than those inoculated only with S. cerevisiae. However, no significant differences were found in alcohol content. The addition of S. cerevisiae tended to standardise the wines and only those made with T. delbrueckii and L. thermotolerans, both alone and together, showed different aromatic profiles. Wines elaborated with non-Saccharomyces yeasts that quickly decreased in tanks showed characteristics similar than those made only with S. cerevisiae. Therefore, sequential inoculation of non-Saccharomyces/Saccharomyces is a useful tool to modulate wine characteristics, but only with species which remain longer in tanks. These findings can be useful to carry out selection processes within these species.


Pediatric urolithiasis in Tunisia: a multi-centric study of 525 patients.
To investigate retrospectively the clinical and epidemiological characteristics, and method of treatment of childhood urolithiasis, a major urological problem in Tunisia. The records of 525 children with urolithiasis treated in Tunisia between 1990 and 2004 were reviewed in a multi-centric study with regard to age at diagnosis, sex, history, and physical, laboratory, and radiologic findings. Metabolic evaluation when performed included serum electrolytes, calcium, phosphorus, uric acid, 24-h urine collection for calcium and creatinine, and a sodium nitroprusside test for cystine. In all cases urine specimens were sent for culture. The stone was located in the upper tract in 420 (80%) and lower tract in 105 children. Of the urine cultures, 40% were positive. Metabolic investigation was performed in 201 patients and was normal in 170 (84%). Urinary stasis secondary to a urinary tract anomaly that led to the formation of stones was found in 77 patients. Stones were treated by surgery (80%), extracorporeal shock wave lithotripsy (ESWL) (5%) and ureteroscopic extraction (4%), and percutaneous nephrolithotomy was performed in 10 patients (2%). The stone passed spontaneously in 9% of cases. The use of ESWL and endourological methods of treatment for childhood urolithiasis must be generalized, with open surgery being reserved for particular and complex cases.


Surfactant-mediated gene transfer for animal cells.
A commercially available cationic surfactant, dimethyl-dioctadecyl ammonium bromide (DDAB), was used for making lipid vesicles. DDAB easily dissolved in water at 60 °C and formed lipid vesicles at room temperature. The lipid vesicles showed very low cytotoxicity compared with other cationic surfactants. After the lipid vesicles were mixed with plasmid DNA solution, the solution was added to mammalian cells. The addition of a nonionic surfactant (Tween 80) to the cationic lipid vesicles at the weight ratio of 1:1 enhanced transfection efficiency. Adding more or less than the optimal amounts of DNA and lipid vesicles resulted in decreased transfection efficiency. With the optimal amounts of DNA (pCMVβ) and lipid vesicles, about 90-95% of CHO-K1 and BHK-21C13 cells transiently expressed β-galactosidase activity 24 h after transfection. By this procedure, stable transformants around 10(5) cells corresponding to 10% efficiency could be obtained by one batch transfection.


Where are the Stents? A Computerized Tracking System to Eliminate the Forgotten Ureteral Stent.
Ureteral stent placement is the main treatment for ureteral obstruction in contemporary urological practice. Meticulous followup is required to confirm ureteral stent removal. Forgotten ureteral stents create significant patient and physician risk. We assessed the efficacy of an electronic spreadsheet tracking system at a VA medical center urology practice to monitor ureteral stent placement and retrieval. From January 2012 to February 2014 a computer based monitoring system was begun for all stents placed in our practice. An Excel® spreadsheet was used to track ureteral stent insertion and removal. The VA Computerized Patient Record System was used to confirm ureteral stent removal. The spreadsheet included an automatic calculator feature to alert technicians/surgeons when stents remained in place 180 days or longer. In 335 patients a total of 379 stents were placed from January 2012 to February 2014. Of these 379 stents 369 were removed or exchanged in 180 days or less. Ten patients died with stents in place and were counted in the removal group for an overall 100% rate of stents accounted for/removed during the study period. An electronic spreadsheet is an effective method to track ureteral stent placement and removal. Manual data entry, and frequent spreadsheet followup and monitoring contributed to a 100% retrieval rate in this study. Future efforts should focus on automated systems that generate reminders with less emphasis on human data entry and tracking.


Three-to-one analog signal modulation with a single back-bias-controlled reconfigurable transistor.
Reconfigurable field effect transistors are an emerging class of electronic devices, which exploit a structure with multiple independent gates to selectively adjust the charge carrier transport. Here, we propose a new device variant, where not only p-type and n-type operation modes, but also an ambipolar mode can be selected solely by adjusting a single program voltage. It is demonstrated how the unique device reconfigurability of the new variant can be exploited for analog circuit design. The non-linearity of the ambipolar mode can be used for frequency doubling without the generation of additional harmonics. Further, phase shifter and follower circuits are enabled by the n- and p-type modes, respectively. All three functions can be combined to create a 3-to-1 reconfigurable analog signal modulation circuit on a single device enabling wireless communication schemes. Both, the concept as well as the application have been experimentally demonstrated on industrial-scale fully-depleted SOI platform. The special transport physics in those structures has been analyzed by TCAD simulations as well as temperature dependent measurements.


A cross-modal interference effect in grasping objects.
The aim of the present paper was to investigate how the kinematics of a hand reaching toward a visual target would be influenced by haptic and proprioceptive input from an unseen distractor actively grasped in the other, nonreaching hand. The main results were that the amplitude of maximum grip aperture was smaller and the time to maximum grip aperture was earlier when the distractor was smaller than the target. The interference effect from the distractor was similar for both hands as they reached. Furthermore, results from a vibrating-distractor condition for passive tactile input revealed that the interference effects were evident only when the distractor was actively grasped. We suggest that neural processing of proprioceptive and tactile information relevant to distractor size produced the observed interference effects. We also emphasize the importance of active manipulation of the distractor stimulus in eliciting such interference effects.


Evaluation of the Claria sharesource system from the perspectives of patient/caregiver, physician, and nurse in children undergoing automated peritoneal dialysis.
Automated peritoneal dialysis (APD) is increasingly preferred worldwide. By using a software application (Homechoice with Claria sharesource system (CSS)) with a mod-M added to the APD device, details of the home dialysis treatment become visible for PD nurses and physicians, allowing for close supervision. We aimed to evaluate the perceptions of patients/caregivers, PD nurses, and physicians about the advantages and disadvantages of CSS. Three different web-based questionnaires for patients/caregivers, nurses, and physicians were sent to 15 pediatric nephrology centers with more than 1 year of experience with CSS. Respective questionnaires were answered by 30 patients/caregivers, 22 pediatric nephrologists, and 15 PD nurses. Most of the nurses and physicians (87% and 73%) reported that CSS improved patient monitoring. A total of 73% of nurses suggested that CCS is not well known by physicians, while half of them reported reviewing CSS data for all patients every morning. Sixty-eight percent of physicians thought that CSS helps save time for both patients/caregivers and healthcare providers by reducing visits. However, only 20% of patients/caregivers reported reduced hospital visits. A total of 90% of patients/caregivers reported that being under constant monitoring made them feel safe, and 83% stated that the patient's sleep quality improved. A remote monitoring APD system, CSS, can be successfully applied with children for increased adherence to dialysis prescription by giving shared responsibility and may help increase the patient's quality of life. This platform is more commonly used by nurses than physicians. Its potential benefits should be evaluated in further well-designed clinical studies with larger patient groups. A higher resolution version of the Graphical abstract is available as Supplementary information.


Use of copeptin and high-sensitive cardiac troponin T for diagnosis and prognosis in patients with diabetes mellitus and suspected acute myocardial infarction.
Diabetes is a major risk factor for acute myocardial infarction (AMI). Assessment of diabetic patients is challenging due to an often atypical presentation of symptoms. We aimed to evaluate the two novel biomarkers copeptin and high-sensitive cardiac troponin (hs-TnT) for the improvement of early diagnosis and risk-stratification in patients with diabetes and suspected AMI. In this prospective international multicenter study we evaluated 379 patients with diabetes in a cohort of 1991 patients presenting with symptoms suggestive of AMI. The measurement of biomarkers was performed at presentation. Among the 379 diabetic patients, 32.7% had AMI, and in the 1621 patients without diabetes, 18.8% had AMI. The additional use of copeptin improved the diagnostic accuracy provided by conventional troponin alone (AUC 0.86 vs. 0.79, p=0.004). During a median follow-up of 814 days, 49 (13.1%) diabetic patients died. Cumulative 2-year survival rate for patients with copeptin levels below 9 pmol/l was 96.6% compared to 82.8% in patients above that level (p<0.001). The same was observed for hs-TnT with a cutoff level of 14 ng/l (97.7% vs. 82.0%, p<0.001) respective of cTnT with a cutoff level of 10 ng/l (93.5% vs. 75.6%, p<0.001). In multivariate Cox analysis, copeptin, hs-TnT and cTnT were strong and independent predictors of 24-month-mortality. Using the dual marker strategy (copeptin and troponin) identified two groups of high-risk patients where 22.5% of the group with hs-cTnT and copeptin above the cutoff and 28.6% with cTnT and copeptin above the cutoff died. In diabetic patients, copeptin only slightly improves the early diagnosis of AMI provided by hs-cTnT. However, both markers (copeptin and troponin) predict long-term mortality accurately and independently of each other.


Study on the Application Value of PBL Combined with Situational Simulation Teaching Method in Clinical Practice Teaching of Radiology Department.
To study the application value of PBL (Problem-Based Learning) combined with a situational simulation teaching method in clinical practice teaching of radiology. 120 interns who practiced in the radiology department of our hospital from 2020 to 2021 were randomly divided into a study group and a control group. The research group used PBL combined with the scenario simulation teaching method to carry out clinical practice teaching activities. The control group was carried out according to the conventional teaching plan. After 3 months, the teaching effect and satisfaction of the two groups were compared. The imaging report quality control score, intravenous contrast agent score, and practice test score in the study group were higher than those in the control group. The scores of the small class and special class in the research group were higher than those in the control group, and the difference was statistically significant (P < 0.05). The scores of job competency questionnaire in the research group were higher than those in the control group (P < 0.05). The occupational self-efficacy questionnaire score of the research group was higher than that of the control group (P < 0.05). The nursing practice satisfaction score of the research group was higher than that of the control group (P < 0.05). The value of PBL combined with the scenario simulation teaching method in the teaching of radiology clinical practice is significant, which can improve the work ability of interns and make them more satisfied.


Matrix effect and correction by standard addition in quantitative liquid chromatographic-mass spectrometric analysis of diarrhetic shellfish poisoning toxins.
An evaluation of the feasibility of liquid chromatography-mass spectrometry (LC-MS) with atmospheric pressure ionization was made for quantitation of four diarrhetic shellfish poisoning toxins, okadaic acid, dinophysistoxin-1, pectenotoxin-6 and yessotoxin in scallops. When LC-MS was applied to the analysis of scallop extracts, large signal suppressions were observed due to coeluting substances from the column. To compensate for these matrix signal suppressions, the standard addition method was applied. First, the sample was analyzed and then the sample involving the addition of calibration standards is analyzed. Although this method requires two LC-MS runs per analysis, effective correction of quantitative errors was found.


Emergence Delirium in Elderly Patients as a Potential Predictor of Subsequent Postoperative Delirium: A Descriptive Correlational Study.
This study aimed to investigate the incidence of emergence delirium (ED) in elderly patients under general anesthesia and to determine the correlation between ED and delirium at five subsequent postoperative days. This research is a descriptive correlational study. A total of 168 aged patients undergoing elective general anesthesia were recruited from a comprehensive tertiary teaching hospital with 2,400 beds in Southern China from April 2018 to September 2018. The Nursing Delirium Screening Scale was used to assess delirium at 30 and 60 minutes after extubation or on discharge from the postanesthesia care unit. Patients were assessed for delirium at postoperative days one through five using the same method. Patients' demographic information, including cognitive function, were collected. Among the 168 aged patients, 58 suffered from ED (34.5%), including the 79.3% for the 46 patients who experienced postoperative delirium (POD). A positive correlation existed between ED and POD (χ2 = 111.744; P < .01). Logistic regression analysis included seven variables: age, preoperative Mini-Mental State Examination score, underlying diseases, American Society of Anesthesiologists grade, surgery duration, postoperative complications, and the presence of ED. Age and ED were concluded to be independent predictive factors of POD. ED in the first hour after tracheal tube removal is a predictor of delirium at five subsequent postoperative days. Accurate and timely assessment of recovery period can effectively guide the treatment and rehabilitation of POD and maximize prevention of adverse consequences.


Comparison of electrocardiographic findings between the midventricular ballooning form and apical ballooning form of takotsubo cardiomyopathy.
Several reports have recently described the variant form of takotsubo cardiomyopathy exhibiting midventricular ballooning. The purpose of this study was to assess electrocardiographic (ECG) findings on admission in patients with midventricular ballooning. ECG findings are different between the midventricular ballooning form and apical ballooning form of takotsubo cardiomyopathy. We reviewed ECGs on admission in 6 patients with midventricular ballooning and 20 patients with apical ballooning. The sum of ST segment elevation in leads V1 to V3 or in leads V4 to V6 was obtained. The number of leads showing ST segment elevation and/or T wave inversion was also obtained. These ECG findings were compared between the 2 groups. In midventricular ballooning, ECG changes including ST segment elevation and/or T wave inversion were observed frequently in leads V2 and V3, and were not observed in leads II, III, -aVR, aVF, V5, and V6. On the other hand, in apical ballooning, they were found in all leads. They were most common in leads V4 and V5. The sum of ST segment elevation in leads V1 to V3 was similar (2.6 ± 2.0 mm vs 2.7 ± 2.0 mm, P = not significant), and the sum of ST segment elevation in leads V4 to V6 was significantly lower in midventricular ballooning than apical ballooning (0.4 ± 0.8 mm vs 3.5 ± 3.0 mm, P<0.05). The number of leads showing ST segment elevation and/or T wave inversion was significantly lower in midventricular ballooning than apical ballooning (3.2 ± 1.0 leads vs 6.3 ± 2.2 leads, P<0.01). Our data suggested that midventricular ballooning may show limited ECG changes despite broad wall motion abnormality.


Multicentre validation of a modified EUCAST MIC testing method and development of associated epidemiologic cut-off (ECOFF) values for rezafungin.
Rezafungin EUCAST MIC testing has been associated with notable inter-laboratory variation, which prevented ECOFF setting for C. albicans. We assessed in vitro susceptibility and reproducibility for a modified EUCAST methodology and established associated wild-type upper limits (WT-ULs). MICs against 150 clinical Candida isolates (six species), molecularly characterized fks mutants (n = 13), and QC strains (n = 6) were determined at six laboratories according to E.Def 7.3 but using Tween 20 supplemented medium. WT-ULs were determined using the derivatization method, the ECOFFinder programme and visual inspection. Consensus WT-ULs were determined. The laboratory- and species-specific MIC distributions were Gaussian with >99.5% MICs within four 2-fold dilutions except for C. parapsilosis (92.8%). The following consensus WT-UL were determined: C. albicans 0.008 mg/L; C. dubliniensis and C. glabrata 0.016 mg/L; C. krusei and C. tropicalis 0.03 mg/L; and C. parapsilosis 4 mg/L. Adopting these WT-UL, six clinical isolates were non-wild-type, five of which harboured Fks alterations. For 11/13 mutants, all 670 MICs were categorized as non-wild-type whereas MICs for C. glabrata Fks2 D666Y and C. tropicalis Fks1 R656R/G overlapped with the corresponding wild-type distributions. Repeat testing of six reference strains yielded 98.3%-100% of MICs within three 2-fold dilutions except for C. albicans CNM-CL-F8555 (96%) and C. parapsilosis ATCC 22019 (93.3%). The modified EUCAST method significantly improved inter-laboratory variation, identified wild-type populations and allowed perfect separation of wild-type and fks mutants except for two isolates harbouring weak mutations. These consensus WT-UL have been accepted as ECOFFs and will be used for rezafungin breakpoint setting.


Efficacy and Safety of Sildenafil in Men With Sexual Dysfunction and Spinal Cord Injury.
Spinal cord injury (SCI) is estimated to affect approximately 276,000 individuals in the United States. Since 2010, the mean age of individuals at the time of the SCI has been 42 years, with nearly 80% of cases involving men. This means that individuals with SCI generally are young men who typically place a great deal of importance on normal sexual and reproductive function. To assess the effect of sildenafil treatment on erectile function and the frequency of ejaculation in men with SCI. This study was a post hoc analysis of pooled data from two randomized, double-blinded, placebo-controlled, flexible-dose, crossover sildenafil trials conducted in Europe, Australia, and Turkey. Two hundred forty-eight men at least 18 years old with traumatic SCI of at least 6 months' duration, with erectile dysfunction solely attributed to SCI, and in a stable heterosexual relationship were treated sequentially with sildenafil and placebo. Exclusion criteria included taking nitrate therapy, severe cardiac failure, and recent stroke or myocardial infarction. The starting sildenafil dose was 50 mg, taken approximately 1 hour before sexual activity, with subsequent dose adjustment to 100 or 25 mg based on efficacy and safety during treatment. There was a 2-week washout between 6-week treatments. Change from baseline in International Index of Erectile Function question 3 (frequency of penetration), question 4 (maintaining erection after penetration), question 9 (frequency of ejaculation), and erectile function domain scores; intercourse success; and treatment preference. All International Index of Erectile Function outcomes, including achieving and maintaining erections and ejaculation frequency, were statistically significantly greater with sildenafil vs placebo, including the subgroup with complete SCI (P < .01 for all comparisons). The percentage of successful intercourse attempts with sildenafil (53% vs 12%) and preference for sildenafil (96% vs 4%) vs placebo were significant (P < .001), including the subgroup with complete SCI. The most common all-cause adverse events with sildenafil were headache (16.1%) and urinary tract infection (11.6%). Sildenafil significantly improves erections, intercourse success, and ejaculation frequency vs placebo, including in men with complete SCI. Sildenafil is an effective and well-tolerated treatment for sexual dysfunction in men with SCI. The increase in frequency of ejaculation could allow the possibility of having children without medical intervention in this patient population. Ohl DA, Carlsson M, Stecher VJ, Rippon GA. Efficacy and Safety of Sildenafil in Men With Sexual Dysfunction and Spinal Cord Injury. Sex Med Rev 2017;5:521-528.


Prevalence of pressure ulcers in hospitals in Brazil and association with nutritional status--a multicenter, cross-sectional study.
Pressure ulcers (PU) represent a widespread, painful, and expensive health care problem directly associated with increased morbidity, mortality, and length of hospital stay. The aim of this study was to determine the prevalence of PU in hospitalised patients in public and private Brazilian institutions and the ulcers' associations with nutritional status and other risk factors. A multicenter, cross-sectional, quantitative and qualitative study was carried out in hospitals in different geographic regions of Brazil from March 2009 to February 2011. The prevalence and characteristics of PU, the nutritional status and the association between the presence of PU, and the nutritional status and other study variables were evaluated. The association of the presence of PU with the study variables was performed by univariate analyses and multivariate logistic regression models. The final multivariate model was one in which all variables were significant at the 0.05 level. According to the subjective global assessment (SGA), the prevalence of PU was 16.9%, and 52.4% of patients were malnourished. PU and their severity were directly associated with malnutrition (P < 0.05). Patients who are bedridden, who are elderly, who have neurological disorders or cancer, who are staying at a public or private institution, and who are staying at the hospital between 8 d and 15 d had an increased risk of PU (P < 0.05). The prevalence of PU in Brazilian general hospitals is high, and the prevalence of malnutrition is extremely high. Malnourishment is one of the most important risk factors associated with the development and severity of PU in hospitals. Patients who are malnourished are more prone to developing PU.


Fatty Acid Mediators in the Tumor Microenvironment.
Patients with cancer frequently overexpress inflammatory cytokines with an associated neutrophilia both of which may be downregulated by diets with high omega-3 polyunsaturated fatty acids (ω-3 PUFA). The anti-inflammatory activity of dietary ω-3 PUFA has been suggested to have anticancer properties and to improve survival of cancer patients. Currently, the majority of dietary research efforts do not differentiate between obesity and dietary fatty acid consumption as mediators of inflammatory cell expansion and tumor microenvironmental infiltration, initiation, and progression. In this chapter, we discuss the relationships between dietary lipids, inflammation, neoplasia and strategies to regulate these relationships. We posit that dietary composition, notably the ratio of ω-3 vs. ω-6 PUFA, regulates tumor initiation and progression and the frequency and sites of metastasis that, together, impact overall survival (OS). We focus on three broad topics: first, the role of dietary lipids in chronic inflammation and tumor initiation, progression, and regression; second, lipid mediators linking inflammation and cancer; and third, dietary lipid regulation of murine and human tumor initiation, progression, and metastasis.


Metastasis to the colon from bronchogenic carcinoma.
Two cases of primary carcinoma of the lung with clinically demonstrated colon metastases are presented. Such metastases may present with intermittent or continuous colonic obstruction, lower gastrointestinal tract bleeding, or anemia. The metastases may be synchronous or metachronous with respect to diagnosis of the lung lesion. Potential problems in management are discussed and the literature, relating to colonic metastasis from bronchogenic carcinoma, is reviewed.


Cardiac resynchronization therapy in a patient with a failing systemic right ventricle.
Systemic right ventricular dysfunction is 1 of the late complications of the atrial switch operation for transposition of the great arteries. It has been reported that cardiac resynchronization therapy (CRT) for the failing systemic right ventricle (RV) improves symptoms and systolic function. However, patient selection for CRT in congenital heart disease is still challenging because the clinical standard for CRT in these patients is not established. We describe a case of successful implantation of a CRT device for a failing systemic RV aided by multimodality imaging and cardiac hemodynamic studies.


Correction: Cerebral Glioma Grading Using Bayesian Network with Features Extracted from Multiple Modalities of Magnetic Resonance Imaging.
[This corrects the article DOI: 10.1371/journal.pone.0153369.].


Radionuclide Therapy for Bone Metastases: Utility of Scintigraphy and PET Imaging for Treatment Planning.
The skeleton is a common site for cancer metastases. Bone metastases are a major cause of morbidity and mortality and associated with pain, pathologic fractures, spinal cord compression, and decreased survival. Various radionuclides have been used for pain therapy. Recently, an α-emitter has been shown to improve overall survival of patients with bone metastases from castration-resistant prostate cancer and was approved as a therapeutic agent. The aim of this article is to provide an overview regarding state of the art radionuclide therapy options for bone metastases, with focus on the role of PET imaging in therapy planning.


Melanoma-associated retinopathy: high frequency of subclinical findings in patients with melanoma.
Melanoma-associated retinopathy (MAR) is a paraneoplastic syndrome with symptoms of night blindness, light sensations, visual loss, defect in visual fields, and reduced b-waves in the electroretinogram. Patients with MAR often suffer from a sudden onset of ocular symptoms that are believed to result from antibody production against melanoma-associated antigens that cross-react with corresponding epitopes on retinal depolarizing bipolar cells. To correlate the frequency of subclinical symptoms suggestive of MAR in melanoma patients to different stages of disease, patient age, type and thickness of the primary tumour, form of therapy, S-100 level and tumour burden. We analysed 28 patients with melanoma in stages I-IV (according to the American Joint Committee on Cancer tumour classification) for the presence of subclinical MAR symptoms using scotopic electroretinography, static and kinetic perimetry and nyctometry. Seven patients had clinical signs and symptoms consistent with MAR, 18 had some indications, while the remaining three had none. We found no correlation between clinical symptoms and stage of disease, tumour burden or S-100 level, but findings suggestive of MAR were observed more frequently in advanced stages of disease. Subclinical retinal involvement characteristic of MAR appears to be more common than previously suspected in patients with cutaneous malignant melanoma. Our findings in this small cohort seem to indicate that the percentage of patients with symptoms suggestive of MAR is higher in advanced stages of disease. Further clinical studies are required to evaluate if the presence of subclinical symptoms suggestive of MAR is correlated with a worse prognosis and a shortened progression-free and overall survival.


Electrochemical biosensor for miRNA-21 based on gold-platinum bimetallic nanoparticles coated 3-aminopropyltriethoxy silane.
We report an electrochemical biosensor based on gold platinum bimetallic nanoparticles (AuPtBNPs)/3-aminopropyltriethoxy silane (APTS) nanocomposite coated fluorine-doped tin oxide (FTO) as a biosensing platform for hybridization-based detection of miRNA-21. Field Emission-Scanning Electron Microscopy (FE-SEM), Fourier Transform Infrared Spectroscopy (FT-IR) and electrochemical measurements were carried out to ensure the successful construction of the biosensor. The amount of cDNA immobilized on electrode surface and hybridization time required for the miRNA-21 sensing were optimized. The biosensing platform showed detection limit of 0.63 fM with wide linear range i.e. 1 fM-100 nM for miRNA-21 detection. The biosensing strategy demonstrates a good recovery yield from 90.18% to 94.6% in serum samples. It offers good selectivity for its complementary miRNA compared to the non-complementary miRNAs. Other analytical features of the biosensor such as stability, reusability and reproducibility were also tested, providing appropriate results.


CT angiography with gadolinium-based contrast media.
To evaluate the potential use of gadolinium (Gd)-based contrast media, especially that of Gadovist, a 1-molar Gd medium, in computed tomography (CT) and compare our findings with standard iodinated contrast media. Using a live rabbit and an acrylic CT body phantom for comparative CT imaging of Gd- and I-based media. The images were acquired at 80, 100, and 120 kVp, using fixed standard beam filtration. The phantom study used serial dilutions of the Magnevist and Ultravist 300 (2.4-molar I), whereas the animal study used different volumes of Gadovist, Magnevist (0.5 molar Gd), and Ultravist administered intravenously. At 80 kVp for the same injection volumes of Gadovist and Ultravist, the image contrast enhancement of the aorta with Gadovist was 40% lower than that of Ultravist. In the phantom studies, however, for the same kVp settings the CT image contrast was up to fourfold higher for Gd compared with iodine when comparing the same molar concentrations of the two elements in the solutions. These results indicate a potential of Gd-based media for clinical CT angiography and provide incentive for further investigation of this subject.


An unusual intraventricular interthalamic vein: two anatomical case reports.
Neurosurgeons use ventricular veins during an endoscopic third ventriculocisternostomy as landmark to progress in ventricles. In the current literature, there is lack of detailed intraventricular venous anatomy. Majority of those papers treats Monro's foramen venous variations. There are no data of third ventricle venous anatomy and variations in the literature. We reported two cases of unusual interthalamic vein that we need to spare during endoscopy.


Visual evoked potentials after hematopoietic allogeneic stem cell transplantation in childhood.
To study visual pathway pathology detected by visual evoked potentials (VEPs) in patients treated with hematopoietic stem cell transplantation (HSCT) in childhood and to determine the impact of adverse ocular findings, somatic diseases, and conditioning regimens on the VEP results. Ophthalmological assessments including pattern VEPs were performed in 47 of 79 patients at a median age of 15 years (range 3-21 years) in median 6 years (1-17 years) after HSCT. Somatic data were extracted from medical records. Eight patients of 47 (17%) demonstrated pathological VEPs with prolonged latencies bilaterally (n = 3) or unilaterally (n = 5) at their latest VEP test at an age of 12-18 years. A subnormal visual acuity was present in 8/11 eyes with pathological VEPs: one eye had cataract, six eyes had cataract surgery where of two had developed secondary cataracts. One eye had residual retinopathy of prematurity. Pathological VEPs were associated with decreased visual acuity (p = 0.00019) but not linked to gender, malignant diagnosis or conditioning. VEP recordings showed an association with decreased visual acuity but no relationship with irradiation or chemotherapy in the present study. VEP recordings might be of clinical value for children with an unexplained subnormal visual acuity undergoing HSCT.


Evaluation of the protective effect of BCG vaccination by a case-control study in Yaounde, Cameroon.
A case-control study carried out in Yaounde (Cameroon) shows that the protective effect of BCG against pulmonary tuberculosis in the young adult was 66% (relative risk 0.34). This result is not affected by taking into account various factors such as sex, age, socio-economic class and geographical origin.


Heterosis-associated proteome analyses of maize (Zea mays L.) seminal roots by quantitative label-free LC-MS.
Heterosis is the superior performance of heterozygous F1-hybrid plants compared to their homozygous genetically distinct parents. Seminal roots are embryonic roots that play an important role during early maize (Zea mays L.) seedling development. In the present study the most abundant soluble proteins of 2-4cm seminal roots of the reciprocal maize F1-hybrids B73×Mo17 and Mo17×B73 and their parental inbred lines B73 and Mo17 were quantified by label-free LC-MS/MS. In total, 1918 proteins were detected by this shot-gun approach. Among those, 970 were represented by at least two peptides and were further analyzed. Eighty-five proteins displayed non-additive accumulation in at least one hybrid. The functional category protein metabolism was the most abundant class of non-additive proteins represented by 27 proteins. Within this category 16 of 17 non-additively accumulated ribosomal proteins showed high or above high parent expression in seminal roots. These results imply that an increased protein synthesis rate in hybrids might be related to the early manifestation of hybrid vigor in seminal roots. In the present study a shot-gun proteomics approach allowed for the identification of 1917 proteins and analysis of 970 seminal root proteins of maize that were represented by at least 2 peptides. The comparison of proteome complexity of reciprocal hybrids and their parental inbred lines indicates an increased protein synthesis rate in hybrids that may contribute to the early manifestation of heterosis in seminal roots. This article is part of a Special Issue entitled: Translational Plant Proteomics.


The position and current status of radiation therapy after primary systemic therapy in breast cancer: a national survey-based expert consensus statement.
Primary systemic therapy (PST) is changing the role of radiation therapy (RT) in breast cancer. Without randomized studies, the optimal indications for RT after PST and surgery are not clear. The present study provides consensus-based recommendations to clarify the role of RT. Radiation oncologists (n = 82; 77 % response rate) in Spain were surveyed to determine their recommendations for locoregional RT following PST and surgery. Most (98 %) specialists support whole breast irradiation after breast-conserving surgery (BCS), regardless of pathologic response to PST. In T1-T2 and T3-T4 tumours with sentinel node biopsy (SNB) prior to PST, 91 and 56 % of respondents, respectively, recommend irradiating the supraclavicular (level IV) and axillary level III nodes when nodal involvement is detected (9 and 44 % of respondents recommend irradiating these areas by independent of nodal status). If SNB is not available, 57 and 30 % of specialists agreed that the aforementioned nodal regions should be irradiated (33 and 65 % of respondents recommend irradiating these areas by independent of nodal status). Between 58 and 76 % of specialists agreed that nodal levels I and II should be irradiated in cases of insufficient lymphadenectomy or when >75 % of the resected nodes are involved. Agreement is strong regarding the indications for local RT after PST and surgery, but less so for nodal irradiation. All patients who undergo BCS should receive RT, even with complete pathologic response. After mastectomy, RT is recommended in all node-positive stage III cases. Prospective studies will clarify indications for RT in this patient population.


Complementary peptide sequences in partner proteins of the adenovirus capsid.
Peptides quite similar to the signal peptide of secretory proteins were found in adenovirus serotype 2 fibre. This was surprising because this protein does not have this property. Because the N-end fibre is implicated in the penton base-fibre interaction and complementary peptides to the N-sequence fibre were found in penton base protein, these complementary peptides might be involved in the protein-protein recognition and interaction process. So far, known structural data agree with such an hypothesis.


Comparison of maternal abdominal subcutaneous fat thickness and body mass index as markers for pregnancy outcomes: A stratified cohort study.
Obesity in pregnancy is associated with a number of adverse outcomes. The effects of central versus general obesity in pregnancy have not been well established. To compare subcutaneous fat thickness (SFT) with body mass index (BMI) as a marker for pregnancy outcomes. A stratified retrospective cohort study was performed on 1200 pregnancies, selected from a total of 4862 nulliparous, nonsmoking women between 2006 and 2010. SFT was measured on routine ultrasound at 18-22 weeks gestation. BMI and SFT measurements were compared for estimating risks for obesity-related pregnancy outcomes using logistic regression adjusted for maternal age. The median SFT was 18.2 mm (range 6.3-50.9 mm), the median BMI was 23.8 kg/m(2) (range 15.2-52.5), and the correlation between SFT and BMI was 0.53. For every 5 mm increase in SFT and every 5 kg/m(2) increase in BMI, the odds ratios for developing gestational diabetes mellitus were 1.40 (CI 1.22-1.61, P < 0.001) and 1.16 (CI 0.95-1.40, P = 0.1), for caesarean section 1.28 (CI 1.16-1.40, P < 0.001) and 1.16 (CI 1.05-1.28, P = 0.003), large for gestational age 1.28 (CI 1.16-1.47, P = 0.001) and 1.10 (CI 0.95-1.28, P = 0.16) and cumulative adverse obesity-related pregnancy outcomes 1.16 (CI 1.10-1.28, P = 0.002) and 1.05 (CI 0.95-1.16, P = 0.45), respectively. SFT at 18-22 weeks gestation is better than BMI as a marker for obesity-related pregnancy outcomes. As SFT is considered a surrogate measure for visceral fat, these results suggest that central obesity is a stronger risk factor than general adiposity in pregnancy.


Uncoupler resistance in E. coli Tuv and Cuv is due to the exclusion of uncoupler by the outer membrane.
The uncoupler resistant bacterial strains E. coli Tuv and Cuv share the high deoxycholate sensitivity of the parent strain, Doc S. However, both Tuv and Cuv show greater resistance than Doc S to other detergents. Measurement of the periplasmic volume indicates that the outer membrane of Doc S is freely permeable to both TPP+ and hydroxymethylinulin. Tuv and Cuv are able to exclude these compounds. EDTA treatment was necessary prior to measuring membrane potential in Tuv and Cuv. Under conditions where delta phi could be measured, uncouplers acted to dissipate delta phi with equal potency in all strains. Uncoupler resistant proline uptake in Tuv and Cuv was abolished by EDTA treatment. Transduction experiments with phage P1 showed that uncoupler resistance could be transferred from Tuv to Doc S. Such transductants were no longer sensitive to novabiocin. The gene for uncoupler resistance cotransduced with the gene pyrE (82 min). Plating efficiency experiments with P1 suggests that detergent sensitivity in Doc S arises from an rfa (81 min) mutation. This mutation is no longer present in Tuv.


Bridging the gap: basic metabolomics methods for natural product chemistry.
Natural products and their derivatives often have potent physiological activities and therefore play important roles as both frontline treatments for many diseases and as the inspiration for chemically synthesized therapeutics. However, the detection and synthesis of new therapeutic compounds derived from, or inspired by natural compounds has declined in recent years due to the increased difficulty of identifying and isolating novel active compounds. A new strategy is therefore necessary to jumpstart this field of research. Metabolomics, including both targeted and global metabolite profiling strategies, has the potential to be instrumental in this effort since it allows a systematic study of complex mixtures (such as plant extracts) without the need for prior isolation of active ingredients (or mixtures thereof). Here we describe the basic steps for conducting metabolomics experiments and analyzing the results using some of the more commonly used analytical and statistical methodologies.


Biofilm development by potentially pathogenic non-pigmented rapidly growing mycobacteria.
A study to evaluate the biofilm-development ability in three different media (Middlebrook 7H9, sterile tap water and PBS-5% glucose) was performed with 19 collection strains from 15 different species on non-pigmented rapidly growing mycobacteria (NPRGM). A microtiter plate assay was developed to evaluate the percentage of covered surface of the microtiter plate wells in different days from day 1 to day 69. All strains were able to develop biofilm in all the tested media. Middlebrook 7H9 showed the fastest growth, followed by sterile tap water and PBS-5% glucose. A sigmoid growth curve was detected in all the strains both in Middlebrook 7H9 and in sterile tap water. A difference could be detected for Mycobacterium abscessus in tap water, where it showed faster growth than all the other strains. Biofilm development seems to be a property of all the species of NPRGM and it depends on the nutrients present in the medium. The microtiter plate assay described here is a useful tool to evaluate differences in biofilm development among the different species of rapidly growing mycobacteria.


Identification of FAD2 and FAD3 genes in Brassica napus genome and development of allele-specific markers for high oleic and low linolenic acid contents.
Modification of oleic acid (C18:1) and linolenic acid (C18:3) contents in seeds is one of the major goals for quality breeding after removal of erucic acid in oilseed rape (Brassica napus). The fatty acid desaturase genes FAD2 and FAD3 have been shown as the major genes for the control of C18:1 and C18:3 contents. However, the genome structure and locus distributions of the two gene families in amphidiploid B. napus are still not completely understood to date. In the present study, all copies of FAD2 and FAD3 genes in the A- and C-genome of B. napus and its two diploid progenitor species, Brassica rapa and Brassica oleracea, were identified through bioinformatic analysis and extensive molecular cloning. Two FAD2 genes exist in B. rapa and B. oleracea, and four copies of FAD2 genes exist in B. napus. Three and six copies of FAD3 genes were identified in diploid species and amphidiploid species, respectively. The genetic control of high C18:1 and low C18:3 contents in a double haploid population was investigated through mapping of the quantitative trait loci (QTL) for the traits and the molecular cloning of the underlying genes. One major QTL of BnaA.FAD2.a located on A5 chromosome was responsible for the high C18:1 content. A deleted mutation in the BnaA.FAD2.a locus was uncovered, which represented a previously unidentified allele for the high oleic variation in B. napus species. Two major QTLs on A4 and C4 chromosomes were found to be responsible for the low C18:3 content in the DH population as well as in SW Hickory. Furthermore, several single base pair changes in BnaA.FAD3.b and BnaC.FAD3.b were identified to cause the phenotype of low C18:3 content. Based on the results of genetic mapping and identified sequences, allele-specific markers were developed for FAD2 and FAD3 genes. Particularly, single-nucleotide amplified polymorphisms markers for FAD3 alleles were demonstrated to be a reliable type of SNP markers for unambiguous identification of genotypes with different content of C18:3 in amphidiploid B. napus.


Gold(III) π-Allyl Complexes.
Gold(III) π-complexes have been authenticated recently with alkenes, alkynes, and arenes. The key importance of PdII π-allyl complexes in organometallic chemistry (Tsuji-Trost reaction) prompted us to explore gold(III) π-allyl complexes, which have remained elusive so far. The (P,C)AuIII (allyl) and (methallyl) complexes 3 and 3' were readily prepared and isolated as thermally and air-stable solids. Spectroscopic and crystallographic analyses combined with detailed DFT calculations support tight quasi-symmetric η3 -coordination of the allyl moiety. The π-allyl gold(III) complexes are activated towards nucleophilic additions, as substantiated with β-diketo enolates.


Characterization of in vitro stability for two processive endoglucanases as exogenous fibre biocatalysts in pig nutrition.
Development of highly efficacious exogenous fibre degradation enzymes can enhance efficiency of dietary fibre utilization and sustainability of global pork production. The objectives of this study were to investigate in vitro stability for two processive endoglucanases, referred to as GH5-tCel5A1 and GH5-p4818Cel5_2A that were overexpressed in CLEARCOLIBL21(DE3). Three-dimensional models predicted presence of Cys residues on the catalytic site surfaces of GH5-tCel5A1 and GH5-p4818Cel5_2A; and time course experimental results shown that both cellulases were susceptible to auto-oxidation by airborne O2 and were unstable. Furthermore, we examined these endoglucanases' stability under the mimicked in vitro porcine gastric and the small intestinal pH and proteases' conditions. Eadie-Hofstee inhibition kinetic analyses showed that GH5-tCel5A1 and GH5-p4818Cel5_2A respectively lost 18 and 68% of their initial activities after 2-h incubations under the gastric conditions and then lost more than 90% of their initial activities after 2-3 h of incubations under the small intestinal conditions. Therefore, further enzyme protein engineering to improve resistance and alternatively post-fermentation enzyme processing such as coating to bypass the gastric-small intestinal environment will be required to enable these two processive endoglucanases as efficacious exogenous fibre enzymes in pig nutrition application.


The relationship of molted primaries of commercial layers to first egg after molt.
The number of primary feathers molted was observed when hens were force molted in two separate experiments. The number of days required for the hens to return to production was directly related (r = .98, Experiment 2) to the number of primary feathers molted.


Reasons of carcinogenesis indicate a big-bang inside: a hypothesis for the aberration of DNA methylation.
Cancer involves various sets of altered gene functions which embrace all the three basic mechanisms of regulation of gene expression. However, no common mechanism is inferred till date for this versatile disease and thus no full proof remedy can be offered. Here we show that the basic mechanisms are interlinked and indicate towards one of those mechanisms as being the superior one; the methylation of cytosines in specific DNA sequences, for the initiation and maintenance of carcinogenesis. The analyses of the previous reports and the nucleotide sequences of the DNA methyltransferases strongly support the assumption that the mutation(s) in the DNA-binding site(s) of DNA-methyltransferases acts as a master regulator; though it continues the cycle from mutation to repair to methylation. We anticipate that our hypothesis will start a line of study for the proposal of a treatment regime for cancers by introducing wild type methyltransferases in the diseased cells and/or germ cells, and/or by targeting ligands to the altered binding domain(s) where a mutation in the concerned enzyme(s) is seen.


Multiple parameter evidence synthesis--a potential solution for when information on drug use and harm is in conflict.
The quality of the evidence on estimating drug-related harm is not yet as advanced as that for intervention effectiveness. Multiple parameter evidence synthesis offers a potential solution, in which 'all available evidence' is combined into a single coherent model.We present a case study of estimating the number of people infected with hepatitis C.


Analysis of specificity determinants in the interactions of different HPV E6 proteins with their PDZ domain-containing substrates.
The E6 oncoproteins of the cancer-associated human papillomaviruses (high-risk HPV types) characteristically have a PDZ-binding motif at their extreme carboxy-termini. However, they interact with only some of the PDZ domain-containing proteins in the human proteome and, despite many of these proteins having multiple PDZ domains, they interact specifically through only one of those domains. Previous work has shown that the exact sequence of the C-terminal PDZ-binding motif of E6 affects substrate selection, and recently we have shown that an E6 residue peripheral to the binding motif also contributes to the specificity of binding. Here we show that substrate specificity of the HPV E6 PDZ binding is modulated both by the amino acid residues upstream of the binding domain and by the non-canonical residues within it. Using this data we have begun to construct a scheme of substrate preferences for E6 proteins from different HPV types.


Antimycobacterial hsp65 and rheumatoid factor titres in a population of normal twins: evidence of genetic control of rheumatoid factor.
Rheumatoid arthritis (RA) is an autoimmune disease and rheumatoid factor (RF), anti-IgG, has been implicated in the pathogenesis, but the exact etiology remains unclear. There are data to suggest and infectious trigger to the autoimmune process, and mycobacteria are considered a candidate. Immunization of various animals with mycobacterial heat shock protein 65 (mhsp65) protects against subsequent autoimmune arthritis in a number of experimental models. Elevated anti-mhsp65 titres have been demonstrated in RA patients, together with specific T cells isolated from inflamed synovium. Mycobacterial hsp65 has also been implicated in other autoimmune disease and in atherosclerosis. The anti-mhsp65 and RF (IgG, IgM and IgA isotypes) titres were assayed by ELISA in 123 pairs of normal twins (61 monozygotic and 62 dizygotic, age 14-79 years), to examine the population distribution and inter-relationship of these antibodies. In addition, we studied the effects of age, sex, genetics and environment on antibody titres. IgG-RF and IgM-RF were detectable in all subjects and IgA-RF in 41 subjects. None of the RF isotypes showed any significant dependence on age or sex. There was a statistically significant correlation between twins for the IgG-RF and IgM-RF, and a positive but not significant correlation for the IgA-RF. All three correlations were stronger for monozygotic than dizygotic twins, reaching statistical significance for IgM-RF (P < 0.001), and this indicates that there is a genetic influence on RF titres. Anti-mhsp65 titres were detectable in 90.5% of the study group with a range of 0.15-19.7 AU/ml. There were weak correlations between twins, stronger for dizygotic than monozygotic twins. This suggests that familial influences on anti-mhsp65 titres are very small, with no evidence of any genetic influence at all. There was no significant relationship of anti-mhsp65 titre with age, sex or RF titres.


Tomographic reconstruction from energy-filtered images of thick biological sections.
Energy filtration makes it possible to image an approximately 0.5 microm biological section at 80 kV in the electron microscope. Based on spectra taken at different tilt angles, we chose the most probable energy loss, deltaEp +/- 10eV for each tilt angle, as the imaging energy window. A complete tilt series from +60 degrees to -60 degrees at 2.5 degree intervals was collected on the Zeiss EM902 and used in a tomographic reconstruction of transcriptionally active chromatin in the Balbiani ring of Chironomus tentans.


Fast Biological Modeling for Voxel-based Heavy Ion Treatment Planning Using the Mechanistic Repair-Misrepair-Fixation Model and Nuclear Fragment Spectra.
The physical and biological differences between heavy ions and photons have not been fully exploited and could improve treatment outcomes. In carbon ion therapy, treatment planning must account for physical properties, such as the absorbed dose and nuclear fragmentation, and for differences in the relative biological effectiveness (RBE) of ions compared with photons. We combined the mechanistic repair-misrepair-fixation (RMF) model with Monte Carlo-generated fragmentation spectra for biological optimization of carbon ion treatment plans. Relative changes in double-strand break yields and radiosensitivity parameters with particle type and energy were determined using the independently benchmarked Monte Carlo damage simulation and the RMF model to estimate the RBE values for primary carbon ions and secondary fragments. Depth-dependent energy spectra were generated with the Monte Carlo code FLUKA for clinically relevant initial carbon ion energies. The predicted trends in RBE were compared with the published experimental data. Biological optimization for carbon ions was implemented in a 3-dimensional research treatment planning tool. We compared the RBE and RBE-weighted dose (RWD) distributions of different carbon ion treatment scenarios with and without nuclear fragments. The inclusion of fragments in the simulations led to smaller RBE predictions. A validation of RMF against measured cell survival data reported in published studies showed reasonable agreement. We calculated and optimized the RWD distributions on patient data and compared the RMF predictions with those from other biological models. The RBE values in an astrocytoma tumor ranged from 2.2 to 4.9 (mean 2.8) for a RWD of 3 Gy(RBE) assuming (α/β)X = 2 Gy. These studies provide new information to quantify and assess uncertainties in the clinically relevant RBE values for carbon ion therapy based on biophysical mechanisms. We present results from the first biological optimization of carbon ion radiation therapy beams on patient data using a combined RMF and Monte Carlo damage simulation modeling approach. The presented method is advantageous for fast biological optimization.


Embolic infarct presented with bilateral thalamic lesions possibly initiated by an emboli to the artery of Percheron.
Bilateral thalamic infarcts are rare and present with varying symptoms. Cardioembolism and small artery disease are the most common etiologic factors. The occlusion of a rare arterial variant called the artery of Percheron results in bilateral thalamic infarcts. Herein, we present the imaging findings of an embolic infarct starting from bilateral thalami, probably due to an emboli to the artery of Percheron first.


The role in cancer therapy of inhibiting recovery from PLD induced by radiation or bleomycin.
Effects on survival of allowing and inhibiting potentially lethal damage recovery (PLDR) after X ray or bleomycin (bleo) exposure, especially at clinically applicable doses, were examined in plateau phase Chinese hamster ovary (CHO) cells. Dose modifying factors (DMF's) between immediately explanted cells and those trypsinized 24 hours after various doses of X rays were 2.51 +/- 0.12 (mean +/- 2 S.E.) at 70% surviving fraction (SF), 2.03 +/- 0.085 at 40% and 1.71-1.79 at lower SF levels. Thus, at doses used clinically (1-4 Gy), the DMF by PLDR was more than 2.0, suggesting its importance in radiotherapy of cancers. Feeder layers were found to increase the SF, especially for replating immediately after bleo exposure. Among the inhibitors tested, 3'-deoxyguanosine at 3.7 mM and caffeine at 10.3 mM inhibited x and bleo PLDR (DMF of 1.0-1.2) when trypsinized 24 hours after treatment. However, interestingly, 3 aminobenzamide, a specific inhibitor of poly (ADP) ribose synthesis, even at 14.7 or 29.4 mM, was not as effective in suppressing both x and bleo PLDR, suggesting the role of repair processes independent of poly (ADP) ribose levels in PLDR in plateau phase cultures. Possibilities of clinical application of PLDR inhibitors as radio- and chemosensitizers are discussed.


Azide-selective sensor based on tripodal iron complex for direct azide determination in aqueous samples.
A potentiometric azide-selective sensor based on the use of iron(III) hydrotris(3,5-dimethylpyrazolyl)borate acetylacetonate chloride [Tp(Me2)Fe(acac)Cl] as a neutral carrier for an azide-selective electrode is reported. Effect of various plasticizers, viz. o-nitrophenyloctyl ether (o-NPOE), dioctylphthalate (DOP), dibutylphthalate (DBP), and benzylacetate (BA), and an anion excluder, hexadecyltrimethylammonium bromide (HTAB), with [Tp(Me2)Fe(acac)Cl] complex in poly(vinyl chloride) (PVC) were studied. The best performance was obtained with a membrane composition of [Tp(Me2)Fe(acac)Cl]/HTAB/DOP/PVC in a ratio of 5:2:190:100 (w/w). The sensor exhibits significantly enhanced selectivity toward azide ions over the concentration range 6.3 x 10(-7) to 1.0 x 10(-2) M with a lower detection limit of 3.8 x 10(-7) M and a Nernstian slope of 59.4 +/- 1.1 mV decade(-1). Influences of the membrane composition, pH and possible interfering anions were investigated on the response properties of the electrode. Fast and stable response, good reproducibility, long-term stability and applicability over a wide pH range (3.5-9.0) are demonstrated. The sensor has a response time of 14 s and can be used for at least 45 days without any considerable divergence in the potential response. The proposed electrode shows fairly good discrimination of azide from several inorganic and organic anions. It was successfully applied to the direct determination of azide in orange juice, tea extracts and human urine samples.


Do Mexican immigrants substitute health care in Mexico for health insurance in the United States? The role of distance.
Although language and culture are important contributors to uninsurance among immigrants, one important contributor may have been overlooked - the ability of immigrants to return to their home country for health care. This paper examines the extent to which uninsurance (private insurance and Medicaid) is related to the ability of immigrants to return to Mexico for health care, as measured by spatial proximity. The data for this study are from the Mexican Migration Project. After controlling for household income, acculturation and demographic characteristics, arc distance to the place of origin plays a role in explaining uninsurance rates. Distance within Mexico is quite important, indicating that immigrants from the South of Mexico are more likely to seek care in their communities of origin (hometowns).


A practical and cost-effective mutation scanning-based approach for investigating genetic variation in Cryptosporidium.
In the present study, we used a mutation scanning-targeted sequencing approach to assess variation in part (pgp60) of the 60 kDa glycoprotein (gp60) gene among Cryptosporidium samples from humans in Victoria, Australia. Two nuclear ribosomal loci (the small subunit rRNA gene and the second internal transcribed spacer) were used to identify the samples as Cryptosporidium hominis (n = 74), Cryptosporidium parvum (n = 23) or Cryptosporidium meleagridis (n = 1). In total, nine distinct pgp60 sequences were identified (three C. hominis, five C. parvum and one C. meleagridis). Phylogenetic analyses of the pgp60 sequence data, employing well-defined reference sequences for comparison, allowed the genotypic and subgenotypic classification of samples. The C. hominis samples were classified as Ib A10G2R2, Id A15G1R2, and a new genotype, designated Ib2, was identified subgenotypically as A18G1R4. The C. parvum samples were classified as IIa A18G3R1, IIa A20G3R1, IIa A22G3R1, IIa A23G3R1 and IIc A5G3R2. These findings suggested that the C. hominis metapopulation is largely homogeneous, consisting of a single dominant genotype, Ib A10G2R2, whereas the C. parvum metapopulation is considerably more heterogeneous, with no single dominant genotype. The greater level of genetic heterogeneity found among the C. parvum samples, despite the smaller sample size, may relate to the zoonotic infection pattern of this species, which would be reflective of a greater number of possible infection sources. The present mutation scanning approach, coupled with targeted sequencing of genetically distinct representatives, is a practical, cost-effective tool for large-scale population genetic and epidemiological studies of Cryptosporidium and other eukaryotic organisms.


Systematic and state-of the science review of the role of environmental factors in Amyotrophic Lateral Sclerosis (ALS) or Lou Gehrig's Disease.
The etiology of sporadic amyotrophic lateral sclerosis (ALS) is still unclear. We evaluate environmental factors suspected to be associated with ALS for their potential linkage to disease causality and to model geographic distributions of susceptible populations and expected cases worldwide. A PRISMA systematic literature review was performed 2021. Bradford Hill criteria were used to identify and rank environmental factors and a secondary review of ALS diagnoses in population studies and ALS case or cohort studies was conducted. Prevalence rate projection informed estimates of impacted regions and populations. Among 1710 papers identified, 258 met the inclusion criteria, of which 173 responded to at least one of nine Bradford Hill criteria among 83 literature-identified ALS environmental factors. Environmental determinants of ALS in order of decreasing significance were β-N-methylamino-L-alanine (BMAA), formaldehyde, selenium, and heavy metals including manganese, mercury, zinc, and copper. Murine animal models were the most common methodology for exploring environmental factors. Another line of investigation of 62 population exposure studies implicated the same group of environmental agents (mean odds ratios): BMAA (2.32), formaldehyde (1.54), heavy metals (2.99), manganese (3.85), mercury (2.74), and zinc (2.78). An age-adjusted incidence model estimated current total ALS cases globally at ~85,000 people compared to only ~1600 cases projected from the reported ALS incidence in the literature. Modeling with the prevalence microscope equation forecasted an increase in U.S. ALS cases from 16,707 confirmed in 2015 to ~22,650 projected for 2040. Two orthogonal methods employed implicate BMAA, formaldehyde, manganese, mercury, and zinc as environmental factors with strong ALS associations. ALS cases likely are significantly underreported globally, and high vulnerability exists in regions with large aging populations. Recent studies on other diseases with environmental determinants suggest the need to consider additional potential triggers and mechanisms, including exposures to microbial agents and epigenetic modifications.


Glycine betaine is the main organic osmotic solute in a stratified microbial community in a hypersaline evaporitic gypsum crust.
We have examined the organic osmotic solutes content within the stratified microbial communities in an evaporitic gypsum crust found in an evaporation pond (~194 g/l total dissolved salts) of the salterns of the Israel Salt Company, Eilat. We extracted intracellular solutes from the upper three pigmented layers of the crust: a yellow-orange layer dominated by unicellular cyanobacteria, a green layer with filamentous cyanobacteria, and a layer colored red-purple by purple sulfur bacteria; dense communities of heterotrophic bacteria were present in all layers. The solutes were analyzed by Raman spectroscopy, (1)H and (13)C nuclear magnetic resonance, and HPLC. All layers contained glycine betaine as the only detectable osmotic solute; ectoine and other solutes known to be produced by many halophilic and halotolerant prokaryotes were not found. In this first attempt to assess the osmotic solute content within complex natural communities of halophilic microorganisms, the predominant role of glycine betaine as an osmolyte was established. Most heterotrophic bacteria cannot produce glycine betaine but preferentially use it when it is supplied. Presence of glycine betaine produced by the photoautotrophic members of the community, therefore, may relieve the heterotrophs from the need to synthesize other compounds at a high-energy cost.


Predictive value of angiographic scores for the integrated management of the ischemic diabetic foot.
To retrospectively evaluate the agreement between the angiographic scores and the clinical outcomes after endoluminal revascularization in diabetic patients with Fontaine stage IV critical limb ischemia (CLI). Clinical and procedural data were retrospectively collected of consecutive diabetic patients with Fontaine stage IV CLI who underwent percutaneous lower limb endoluminal revascularization from January 2009 to June 2011. Pre- and postprocedural angiographic images were retrospectively reviewed to classify lower limb arterial involvement according to four systems: (1) TransAtlantic Inter-Society Consensus [TASC] I; (2) TASC II; (3) Graziani's morphologic classification; and (4) Joint Vascular Society Council calf and foot scores. Foot lesions were graded according to the University of Texas wound classification system. Clinical results (healing, nonhealing, or major amputation) were compared with baseline clinical data and angiographic results. In the study period, 202 percutaneous procedures were performed, with an immediate technical success rate of 94%. Preprocedurally, the mean ± standard deviation calf and foot scores were 7.8 ± 1.6 and 7.3 ± 2.3, respectively; 132 patients (65%) were in Graziani's morphologic classes from 4 to 7; in 112 (55%) cases, TASC II was considered inapplicable, for the absence of femoropopliteal lesions; and finally, 93% of limbs were classified as TASC I type D lesions. After the procedure, mean calf and foot scores were 4.8 ± 2.3 and 5.9 ± 2.6, respectively, and 87% of cases were in Graziani's classes 1 and 2; TASC II was inapplicable in all cases, whereas 80% of cases remained TASC I type D lesions. Healing rate was 67% and major amputation rate was 4%. Among all the clinical and angiographic variables included in the analysis, only pre- and postprocedural foot scores were significantly associated to the clinical outcome (P < .05). Endoluminal revascularization represents a valuable treatment option in diabetic patients with CLI. TASC classifications are inadequate to describe peripheral arterial involvement in the vast majority of diabetic patients with CLI. Pre- and postprocedural foot scores represent the most significant angiographic parameters to evaluate treatment success.


Potabilization of low NOM reservoir water by ultrafiltration spiral wound membranes.
Membrane technologies such as ultrafiltration offer an interesting alternative to integral treatment of surface water destined for human consumption. With this in mind, a pilot-scale ultrafiltration module was set up, equipped with spiral-wound polyethersulphone membranes (16.6m(2)) with an effective pore size of 0.05 microm. The system operated continuously with a stable production of 0.9 m(3)/h (54 lmh) and a constant transmembrane pressure of -0.2 bar. The effluent obtained showed a total absence of faecal contamination indicators of both bacterial and viral origin, and also presented an excellent physico-chemical quality, independently of the quality of influent. Total aerobic bacteria counts revealed the problem of bacterial contamination in the membrane permeate zone, which could be controlled through daily chemical cleansing of the membrane. The chief problem presented by this type of system, applied as exclusive treatment, is low effectiveness in the retention of natural organic matter (NOM), in which respect the quality of the effluent was observed to depend on the quality of influent. This constitutes the principal limitation for applying the system to surface water due to the risk of disinfection by-products formation during the final post-chlorination. However, spiral wound ultrafiltration (SWUF) membranes could be used for low NOM reservoir water total treatment offering several advantages over conventional technologies.


Clinical outcomes of the use of unidirectional barbed sutures in gastrointestinal surgery for dogs and cats: A retrospective study.
To report the clinical outcomes of gastrointestinal surgery using unidirectional barbed sutures in single-layer appositional closure in dogs and cats. Retrospective and descriptive study. Twenty-six client-owned dogs; three client-owned cats. Medical records of dogs and cats that received gastrointestinal surgery closed with unidirectional barbed sutures were reviewed to collect information on signalment, physical examinations, diagnostics, surgical procedures, and complications. Short- and long-term follow-up information was collected from the medical records, the owners, or the referring veterinarians. Six gastrotomies, 21 enterotomies, and nine enterectomies were closed with a simple continuous pattern with unidirectional barbed glycomer 631 sutures. Nine dogs had multiple surgical sites closed with unidirectional barbed sutures. None of the cases in the study developed leakage, dehiscence, or septic peritonitis during the 14-day short-term follow up. Long-term follow up information was collected for 19 patients. The median long-term follow-up time was 1076 days (range: 20-2179 days). Two dogs had intestinal obstruction due to strictures at the surgical site 20 and 27 days after surgery. Both were resolved with an enterectomy of the original surgical site. Unidirectional barbed suture was not associated with a risk of leakage or dehiscence after gastrointestinal surgery in dogs and cats. However, strictures may develop in the long term. Unidirectional barbed sutures can be used during gastrointestinal surgery in client-owned dogs and cats. Further investigation of the role of unidirectional barbed sutures leading to abscess, fibrosis, or stricture is necessary.


Male skin and ingredients relevant to male skin care.
Male skin care needs are heavily influenced by the need to remove facial hair on a regular basis. Facial skin issues associated with poor hair removal approaches are common and include razor burn and irritation. This paper evaluates current research on shaving technology and how careful ingredient selection can contribute to male skin health. The importance of maintaining hair softness during the shave and restoring facial hydration post-shave is discussed. Data are presented on how post-shave moisturizers containing glycerine and emollients can create an environment for improved barrier function which can be further improved by incorporating specific ingredients such as niacinamide.


Luminance discrimination, color contrast, and multiple mechanisms.
The purpose of this study was to investigate the effect of color contrast on luminance discrimination. Observers were required to indicate the more intense of two stimuli presented briefly in a surround. In some conditions the two stimuli were the same chromaticity as the surround, while in other conditions the stimuli differed in chromaticity from the surround. The luminance of the fixed test stimulus was varied in different conditions over a range from below the surround level to above the surround level. Difference thresholds were proportional to the luminance difference between test and surround over much of the range. However, difference thresholds were higher at low luminance contrasts when the chromatic contrast between the stimuli and the surround was high. Results also indicate that the effects of chromatic contrast may be mediated by local contrast mechanisms, but that the relationship between threshold and luminance contrast is not mediated entirely by these local contrast mechanisms.


There is an inverse relationship between the capacity of climate change refugia and species adaptation potential.
There is an inverse relationship between species adaptation potential (green/curved line) and refugial capacity (blue/light triangle). The ability of refugia species to adapt/acclimatize to future stressors (red/dark triangle), assuming this is within their physiological capabilities, depends on the refugial capacity of the refugia in which they live. The more effective a refugium, the lower the chance of its species adapting/acclimatizing. This article is a commentary on Kapsensberg & Cyronak, 25, 3201-3214.


Sustainable spectrophotometric determination of antihypertensive medicines reducing COVID-19 risk via paired wavelength data processing technique - Assessment of purity, greenness and whiteness.
Recent studies have reported that using certain antihypertensive therapies such as angiotensin II receptor blockers (ARBs) is associated with mitigation of fatal outcomes and enhancing clinical features of patients having hypertension during coronavirus pandemic. Thus, in the current work an innovative, effective, white and sustainable spectrophotometric technique called paired wavelength data processing technique (PWDPT) was developed for evaluation of recommended antihypertensive combination therapies incorporating candesartan cilexetil (CAN) and hydrochlorothiazide (HCT). This technique included three methods, namely, absorbance resolution (AR), amplitude resolution (PR) and ratio extraction (RE). Linearity ranges were (5.0 μg/mL - 50.0 μg/mL) and (2.0 μg/mL - 24.0 μg/mL) for CAN and HCT, respectively. Validation and confirmation of all suggested methods were conducted in accordance with ICH guidelines, producing satisfactory results within the accepted limits. Statistical comparison was achieved between the attained results from suggested methods and those attained from official methods, in which insignificant difference was existed. The suggested methods were successfully employed for identification of the studied drugs as well as determination of their spectral recognition and evaluation of the purity in their combined formulations. The proposed methods followed the principles of green analytical chemistry, where their greenness was evaluated and compared with the official potentiometric and HPLC methods via using four tools, namely, National Environmental Methods Index (NEMI), the Analytical Eco-Scale, the Green Analytical Procedure Index (GAPI) and Analytical greenness metric (AGREE) which affirmed the eco-friendly nature of the proposed methods. Moreover, studying the whiteness features was performed using the recently introduced RGB12 model. The acceptable results along with the sustainability, simplicity, affordability and low-cost of the proposed methods encourages their utilization in the quality control laboratories.


Properties of after-discharges from cortical electrical stimulation in focal epilepsies.
This study sought to determine whether certain aspects of after-discharges (ADs) obtained during cortical functional mapping provide better correlations between stimulus site and that of spontaneous seizures. Secondly, we wished to determine the percentage of stimulations evoking ADs and, of these, the percentage which clearly involves more than one electrode position, potentially inaccurately localizing cortical function. Thirdly, we wished to quantify the incidences of the several AD morphologies described by Jasper [in: Epilepsy and the functional anatomy of the human brain, 1954, p. 183; p. 692] and to assess whether certain morphologies had a greater tendency to evolve in frequency or morphology. In these 29 patients requiring invasive recordings to determine principal epileptogenic areas, only subdural strips were placed in 19 patients, only grids in 2 patients, and both in 8 patients. A median of 21 electrodes per patient was stimulated of a median of 63 electrodes placed, with the following parameters: biphasic, monopolar, 50 Hz, 0.3 ms pulse duration, 1.5-18 mA. Coverage involved the frontal and parietal lobes (9 patients), frontal parietal temporal lobes (8), frontal temporal (3), temporal (2), occipital (2) and occipital temporal (2) with other combinations in 3 additional patients. Classification of AD morphologies was determined by a pilot study using IFSECN definitions [Electroenceph clin Neurophysiol 1974;37:538] and descriptions by Jasper [in: Epilepsy and the functional anatomy of the human brain, 1954, p. 183] and Gloor [in: Advances in neurology, vol. 8, 1975; p. 59]. Four hundred and two ADs (12%) were elicited by 3358 trains of electrical stimuli of which 260 (65%) clearly involved more than only the stimulated electrode position. Thus, 260 (8%) of 3358 stimulations evoked an AD that could mislocalize cortical function. The proportion of stimulating electrodes eliciting ADs ranged among patients from 4 to 83% (median 33%). Polyspike bursts and sequential spikes were the most common AD morphologies. Ten percent of ADs evolved in morphology, frequency or both. Evolution occurred more commonly (44%) with rhythmic waves than with other AD morphologies (7%). Neither evolving ADs, ADs producing clinical seizures, or ADs exceeding 10 s correlated topologically with spontaneous seizure origins. Although occurring in a minority of cortical electrical stimuli, ADs may involve more than the stimulus site and therefore may inaccurately localize cortical function. Our material failed to disclose any consistent relationship between the site of stimulus eliciting ADs and that of spontaneously appearing seizures, even when certain aspects of such ADs were analysed. These data illustrate the need for scrutiny of the post-stimulus electrocorticogram for ADs and particularly those involving extra stimulus sites. Whether varying the stimulus parameters prospectively will disclose a better seizure localizing value for ADs remains to be determined.


Candidate genes and potential targets for therapeutics in Wilms' tumour.
Wilms' tumour (WT) is the most common malignant renal tumour of childhood. During the past two decades or so, molecular studies carried out on biopsy specimens and tumour-derived cell lines have identified a multitude of chromosomal and epigenetic alterations in WT. In addition, a significant amount of evidence has been gathered to identify the genes and signalling pathways that play a defining role in its genesis, growth, survival and treatment responsiveness. As such, these molecules and mechanisms constitute potential targets for novel therapeutic strategies for refractory WT. In this report we aim to review some of the many candidate genes and intersecting pathways that underlie the complexities of WT biology.


A new model validation tool using kernel regression and density estimation.
In physiological system modelling for control or decision support, model validation is a critical element. A nonparametric approach for assessing the validity of deterministic dynamic models against empirical data is developed, based on kernel regression and kernel density estimation, yielding visual graphical assessment tools as well as numerical metrics of compatibility between the model and the data. Nonparametric regression has been suggested for assessing a parametric statistical model by constructing a confidence band for the proposed model and then checking whether the nonparametric regression curve lies within the band. However, for deterministic models, there is no confidence band that can be constructed. A reversal of roles is therefore suggested--construct a probability band for the nonparametric regression curve and check whether the proposed model lies within the band. This approach extends the utility of nonparametric regression for model assessment to deterministic models. Weighted kernel density estimation is incorporated to derive a density profile for the regression curve, creating a local graphical validation tool. In addition, the density profile is used to define and compute two numerical measures--average normalized density (AND) and relative average normalized density (RAND), representing global statistical validity measures. These tools are demonstrated using a biomedical system model for agitation-sedation and sedation management control.


A static and dynamic NMR study of 10-hydrazino-BODIPY.
10-Hydrazino-BODIPY, BoNHNH2 , presents slow rotation about the C10-NH bond that results in anisochronous 1 H and 13 C NMR signals. The assignment of the different signals has been made using traditional two-dimensional methods as well as spin-spin coupling constants and confirmed by DFT calculations (B3LYP) using the 6-311++G(d,p) basis set. The rotational barrier has been determined in three pairs of proton signals and compared with the calculated barrier.


Sex Differences and Emerging New Risk Factors for Atherosclerosis and Its Thrombotic Complications.
Cardiovascular diseases (CVD) remain the world's leading cause of death and disability in both men and women, but with different prognostics and outcomes between sexes. Although the burden of CVD is generally related to the conventional risk factors, the relevance of non-traditional risk factors is increasingly being recognized to explain the so-called "residual risk". Men and women share many similarities regarding classical cardiovascular risk factors but have different disease pathophysiology, clinical presentations, prevalence, and outcomes of CVDs. How sex-specificities regarding the effects of non-traditional risk factors may contribute to the evolution of atherosclerosis and its clinical manifestations in males and females remain largely underanalyzed. The present review summarizes the current knowledge for sex differences in atherosclerotic plaque composition and clinical evolution in association with risk factors, such as inflammation, lipoprotein(a), hemostasis, intraplaque calcification, and depression. We further discuss the potential sex-differential impact of chronic infectious diseases, gut microbiome and, epigenetic gene expression regulation for atherosclerosis and the effect of female-specific disorders in CVD.


Type 1 (11;22)(q24:q12) translocation is common in Ewing's sarcoma/peripheral neuroectodermal tumour in south Indian patients.
The Ewing's sarcoma family can present diagnostic difficulties. In the past the basis of diagnosis has been a exclusion. Identification of a specific translocation especially t(11;22) (EWS-FLI 1 fusion gene), which is seen in nearly 85 percent of Ewing's sarcoma cases can help in precise diagnosis. We have carried out a study on twenty patient samples diagnosed to have Ewing's sarcoma/peripheral neuroectodermal tumour (PNET)/small round cell malignant tumour. The study involved RT-PCR analysis for the fusion transcript, followed by sequencing to identify the specific type of fusion. Ninety percent (18/20) of the samples tested were found to be t(11;22) translocations involving EWS-FLI 1 genes. Sixty-one percent (11/18) were found to be type 1 fusion and seven were type 2 (39 percentage). This is the first study in India with quantitative information about the types of EWS-FLI 1 translocations present in Ewing's family of tumours in south Indian patients.


An atypical lipomatous tumor of the hypopharynx, a unique entity? A rare case report.
Liposarcomas are thought to be the most prevalent kind of soft tissue sarcoma in adults. A well-differentiated liposarcoma that has a higher incidence of a local recurrence following surgical excision is referred to as an atypical lipomatous tumor. The incidence are extremely rare in which <1 % of head and neck sarcoma cases. This is unusual liposarcoma localization warrants great interest in reporting such a case. In this report we present a case of a 50-year-old male complained with difficulty swallowing solid food and continuous presence of lump in the throat. Fiber Optic Laryngoscopy (FOL) showed a tumor filled the hypopharynx and CT scan suggested a benign mass that was most likely a fibrolipoma. Tumor was found infiltrating the lateral pharyngeal wall and protruding into the hypopharyngeal lumen. Because the tumor had spread to the right thyroid lobe, surgical excision via a transcervical approach was combined with a right thyroidectomy. There was a positive margin at the end of resection, thus a chemoradiation was added. Two years post-operative evaluation shows no sign of recurrence. The main treatment for hypopharyngeal liposarcoma is based on surgery, either through an endoscopic or transcervical approach which determined by the size of the tumor and surgical field. Adjuvant chemoradiation is given to help prevent the recurrence.


Redo mitral valve replacement in children.
Despite excellent results with current techniques of mitral valve repair, some children still require mitral valve replacement (MVR). Of necessity, a certain percentage of these children need valve rereplacement (redo MVR). Of 82 MVRs at our institution since 1988, 22 were redo MVRs in 21 children aged 1.4-21 years (mean 9.8 +/- 5.6 years). Interval from initial MVR was 1 month to 18 years (mean 5.8 +/- 4.9 years). Reoperation indication was primarily stenosis in 13 (usually pannus formation), regurgitation in 4, valve thrombosis or embolism in 4, and endocarditis in 1. The initial valve was mechanical in 18 and xenograft in 4. During redo MVR, 8 of 22 (36%) patients had additional procedures including left ventricular outflow tract obstruction relief or aortic valve replacement in 4 and tricuspid valve repair in 2. The new mitral valve was mechanical in 20 and xenograft in 2. Median valve size increased from 21 mm to 25 mm although 7 valves were not upsized on rereplacement (average valve size increase was 3.3 +/- 1 mm). Only 2 patients (9%) needed a new pacemaker. There were no hospital deaths. Six children developed end-stage heart failure and underwent successful cardiac transplantation 3 to 32 (mean 12.1 +/- 10.0) months, postoperatively. On follow-up of 1 month to 7.7 years (mean 2.5 +/- 2.3 years) there has been only one late death of an unrecognized coronary artery anomaly. Redo MVR in children can be performed safely with low morbidity and mortality. A larger prosthesis can often be placed in these children. A surprisingly high percentage of patients eventually require transplantation, perhaps reflecting unnecessary delay in referral for redo MVR.


Applying active learning in a virtual classroom such as a molecular biology escape room.
The transition to remote-teaching online for Molecular Biology has forced active learning exercises like Escape Rooms to also move online. In the past, Escape Rooms have been an effective tool for students to help reinforce concepts they learned in Molecular Biology. We propose that there is a way for Escape Rooms to be moved to an online setting and still be an effective avenue for students to learn the material in a fun and interactive way.


A longitudinal study of the biological variability of plasma lipoproteins in healthy young adults.
The fasting serum concentrations of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total triglyceride and apoprotein A1 were measured at intervals of 12-18 weeks for 60 weeks in 17 male and 11 female healthy young adults in order to assess the variability of these risk factors for coronary disease. No statistically significant seasonal changes were detected in any variable in either sex, although a progressive rise in apoprotein A1 concentration was observed. The coefficients of variation for random fluctuations with time were in the rank order: total cholesterol less than HDL-C less than apoprotein A1 less than LDL-C less than triglyceride. These differences were attributable to biological, rather than to methodological, factors. Within subjects, HDL cholesterol concentration varied inversely with triglyceride concentration and directly with apoprotein A1 concentration. The marked differences which exist in the biological variability of lipid risk factors for atherosclerosis need to be taken into account when making comparisons in epidemiological studies of the predictive powers of single on-entry measurements for future disease. Fluctuations of HDL-C with time appear to be related in part to variations in triglyceride-rich lipoprotein metabolism.


Robust tip localization under continuous spatial and temporal constraints during 2D ultrasound-guided needle puncture.
During ultrasound-guided (US-guided) needle puncture for minimally invasive procedures, automated needle tip localization can help clinicians capture small tips in US images easily and precisely, providing them with obvious tip indicators on the screen and bringing them more confidence during the procedures. However, automated needle tip localization in US images is challenging due to serious interferences arising from all kinds of echoes. We propose a method that localizes needle tips under continuous spatial and temporal constraints in the real-time US frame stream. A temporal constraint is firstly acquired by detecting translational tip motion in motion-enhanced US images with a deep learning-based (DL-based) detector. A spatial constraint and candidate tip locations are obtained by detecting needle shafts and tips in the raw grayscale B-mode images with another DL-based detector. To provide continuous constraints, estimated tip velocity from acquired temporal constraint is used to predict tip locations in frames where no temporal or spatial constraint is detected. Finally, tip coordinates are precisely localized among candidate tips under the spatial and temporal constraints. Experimental results evaluated on 1121 US images from porcine organ punctures, and 895 images from human thyroid punctures demonstrate that the proposed method is effective and efficient, surpassing existing methods. On porcine organ data, a 97.2% recall rate and a 91.9% precision rate on tip detection and 0.88 ± 0.70 mm root-mean-square error (RMSE) on tip localization were achieved. On the human thyroid data, which was not involved in the training, 86.5% recall, 84.3% precision and 0.92 ± 0.78 mm RMSE were achieved separately. The running speed of 14.5 frames per second was achieved only using a CPU. The proposed method provides a more reliable solution for automated needle tip localization during US-guided needle puncture, being more robust to interferences. Fast running speed leads to its practicability in the real-time US stream.


Impaired trafficking and instability of mutant kidney anion exchanger 1 proteins associated with autosomal recessive distal renal tubular acidosis.
Mutations in solute carrier family 4 member 1 (SLC4A1) encoding anion exchanger 1 (AE1) are the most common cause of autosomal recessive distal renal tubular acidosis (AR dRTA) in Southeast Asians. To explain the molecular mechanism of this disease with hematological abnormalities in an affected family, we conducted a genetic analysis of SLC4A1 and studied wild-type and mutant AE1 proteins expressed in human embryonic kidney 293T (HEK293T) cells. SLC4A1 mutations in the patient and family members were analyzed by molecular genetic techniques. Protein structure modeling was initially conducted to evaluate the effects of mutations on the three-dimensional structure of the AE1 protein. The mutant kidney anion exchanger 1 (kAE1) plasmid construct was created to study protein expression, localization, and stability in HEK293T cells. We discovered that the patient who had AR dRTA coexisting with mild hemolytic anemia carried a novel compound heterozygous SLC4A1 mutations containing c.1199_1225del (p.Ala400_Ala408del), resulting in Southeast Asian ovalocytosis (SAO), and c.1331C > A (p.Thr444Asn). Homologous modeling and in silico mutagenesis indicated that these two mutations affected the protein structure in the transmembrane regions of kAE1. We found the wild-type and mutant kAE1 T444N to be localized at the cell surface, whereas the mutants kAE1 SAO and SAO/T444N were intracellularly retained. The half-life of the kAE1 SAO, T444N, and SAO/T444N mutants was shorter than that of the wild-type protein. These results suggest impaired trafficking and instability of kAE1 SAO/T444N as the likely underlying molecular mechanism explaining the pathogenesis of the novel SLC4A1 compound heterozygous mutation identified in this patient.


The dark kinase STK32A regulates hair cell planar polarity opposite of EMX2 in the developing mouse inner ear.
The vestibular maculae of the inner ear contain sensory receptor hair cells that detect linear acceleration and contribute to equilibrioception to coordinate posture and ambulatory movements. These hair cells are divided between two groups, separated by a line of polarity reversal (LPR), with oppositely oriented planar-polarized stereociliary bundles that detect motion in opposite directions. The transcription factor EMX2 is known to establish this planar polarized organization in mouse by regulating the distribution of the transmembrane receptor GPR156 at hair cell boundaries in one group of cells. However, the genes regulated by EMX2 in this context were previously not known. Using mouse as a model, we have identified the serine threonine kinase STK32A as a downstream effector negatively regulated by EMX2. Stk32a is expressed in hair cells on one side of the LPR in a pattern complementary to Emx2 expression in hair cells on the opposite side. Stk32a is necessary to align the intrinsic polarity of the bundle with the core planar cell polarity (PCP) proteins in EMX2-negative regions, and is sufficient to reorient bundles when ectopically expressed in neighboring EMX2-positive regions. We demonstrate that STK32A reinforces LPR formation by regulating the apical localization of GPR156. These observations support a model in which bundle orientation is determined through separate mechanisms in hair cells on opposite sides of the maculae, with EMX2-mediated repression of Stk32a determining the final position of the LPR.


Alteration of the migration trajectory of antibiotic resistance genes by microplastics in a leachate activated sludge system.
The environmental behavior of emerging contaminants of microplastics (MPs), antibiotics and antibiotic resistance genes (ARGs) in the leachate activated sludge system has been monitored and analyzed comprehensively. The results suggested that MPs could effectively alter the migration trajectory of tetracycline resistance genes (tet genes) in the leachate activated sludge system under intermittent and continuous influent conditions. After adding MPs, the total average abundance of tet genes in leachate increased from 0.74 ± 0.07 to 0.78 ± 0.07 (log10tet genes/log10 16S rRNA) and that in sludge increased from 0.65 ± 0.08 to 0.70 ± 0.06 (log10tet genes/log10 16S rRNA). Except for tetA, the abundance of tetB, tetO, tetM and tetQ on MPs increased with increasing TC concentration under both aerobic and anaerobic conditions. MPs not only significantly affect the abundance level and migration trajectory of ARGs in the leachate activated sludge system, but also remarkably improve the level of heavy metals in the ambient environment, indirectly promoting the selective effect of antibiotic-resistant bacteria (ARB) and promoting the development of antibiotic resistance (AR). In addition, MPs changed their physicochemical properties and released hazardous substances with aging to force tet genes to migrate from the leachate activated sludge system to the MPs, making AR more difficult to eliminate and persisted in wastewater treatment plants. Meanwhile, microorganisms played a driving role, making MPs serve as a niche for ARGs and ARB colonization. The co-occurrence network analysis indicated the specific distribution pattern of tet genes and microorganisms in different media, and the potential host was speculated. This study improves the understanding of the environmental behavior of emerging contaminants in leachate activated sludge system and lays a theoretical for protecting the ecological environment.


Establishment of an induced pluripotent stem cell (iPSC) line from a patient with Clozapine-responder Schizophrenia.
Peripheral blood mononuclear cells (PBMCs) were collected from a patient with treatment-refractory Schizophrenia who presented an exceptional clinical response to Clozapine. iPSC lines were established with a non-integrating reprogramming system based on Sendai virus. A footprint-free hiPSC line was characterized to confirm the expression of the main endogenous pluripotency markers and have a regular karyotype. Pluripotency was confirmed by differentiation into cells belonging to the three germ layers. This hiPSC line represents a valuable tool to study the molecular, biochemical and electrophysiological properties of mature neuronal populations belonging to Clozapine responder patients with a severe form of Schizophrenia.