Toxicology and Carcinogenesis Studies of Furosemide (CAS No. 54-31-9) in F344/N Rats and B6C3F1 Mice (Feed Studies).
Furosemide is a diuretic used in human and veterinary medicine. Toxicology and carcinogenesis studies were conducted by feeding diets containing furosemide (99% pure, USP grade) to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, mouse L5178Y lymphoma cells, and Chinese hamster ovary (CHO) cells. Fourteen-Day and Thirteen-Week Studies: Dietary concentrations of furosemide used in the 14-day studies for rats and mice ranged up to 46,000 ppm. Two of five male and 3/5 female rats that received 46,000 ppm furosemide died before the end of the studies. Rats that received 15,300 or 46,000 ppm lost weight over the course of the studies. The final mean body weights of rats that received 1,700 or 5,100 ppm were 12% or 23% lower than that of controls for males and 8% or 16% lower for females. Nephrosis was dose related in rats. All five male and 1/5 female mice that received 46,000 ppm furosemide died before the end of the 14-day studies. Male mice that received 15,300 ppm and female mice that received 46,000 ppm lost weight. The final mean body weights of male mice that received 1,700 or 5,100 ppm were 16% or 14% lower than that of controls. The final mean body weight of females that received 15,300 ppm was 13% lower than that of controls. Slight dilatation of the renal cortical tubules and/or nephrosis were dose related in mice. Dietary concentrations of furosemide used in the 13-week studies were 0 and 625-10,000 ppm for male rats and 0 and 938-15,000 ppm for female rats and male mice. Concentrations for female mice were 0 and 1,250-20,000 ppm. None of the rats died before the end of the studies. The final mean body weights of male rats that received 2,500, 5,000, or 10,000 ppm furosemide were 11%, 22%, or 44% lower than that of controls. The final mean body weights of female rats that received 3,750, 7,500, or 15,000 ppm were 18%, 26%, or 35% lower than that of controls. Minimal-to-mild nephrosis occurred in the two highest dose groups of male and female rats. Mineralization of minimal to mild severity was observed at the renal corticomedullary junction in dosed male rats receiving 625 ppm or more; the severity and incidence of the mineralization increased with increased dose. No compound-related deaths occurred in mice. The final mean body weights of male mice that received 3,750, 7,500, or 15,000 were 12%, 22%, or 17% lower than that of controls. Final mean body weights of dosed and control female mice were comparable. Compound-related lesions in mice induced minimal-to-mild nephrosis. Because of the lower body weights and the kidney lesions in the 13-week studies, doses selected for the 2-year studies were 0, 350, or 700 ppm furosemide in the diet for groups of 50 F344/N rats of each sex. Groups of 50 B6C3F1 mice of each sex were fed diets containing 0, 700, or 1,400 ppm furosemide for 104 weeks. Body Weight and Survival in the Two-Year Studies: Mean body weights of dosed and control rats were comparable throughout the studies. No significant differences in survival were observed between any groups of rats of either sex (final survival--male: control, 17/50; low dose, 17/50; high dose, 20/50; female: 35/50; 31/50; 34/50). The final survival of all groups of male rats was low, reflecting the large number of moribund animals killed after week 91. Survival at week 90 was 35/50, 28/50, and 34/50. Mean body weights of high dose male mice were up to 17% lower than those of controls, and mean body weights of low dose male mice were about 5%-10% lower than those of controls after week 31. Mean body weights of high dose female mice were up to 22% lower than those of controls. Mean body weights of low dose female mice were 5%-13% lower than those of controls after week 82. The survival of the high dose group of female mice was significantly lower than that of controls after week 99 (final survival--male: 31/50; 24/50; 26/50; female: 36/50; 29/50; 18/50). Feed consumption by dosed rats was similar to that by controls. The estimateer week 99 (final survival--male: 31/50; 24/50; 26/50; female: 36/50; 29/50; 18/50). Feed consumption by dosed rats was similar to that by controls. The estimated average amount of furosemide consumed per day was approximately 14-16 or 29-31 mg/kg for low dose or high dose rats. Feed consumption by dosed mice was approximately 5%-7% greater than that by controls. The average amount of furosemide consumed per day was approximately 91-99 or 191-214 mg/kg for low dose or high dose mice. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Nephropathy occurred at similar incidences in all groups of rats, but the severity was greater in dosed male rats. Tubular cell hyperplasia was observed in 4/50 control, 2/50 low dose, and 4/50 high dose male rats. Tubular cell adenomas of the kidney occurred in 1/50 control, 3/50 low dose, and 1/50 high dose male rats. Tubular cell adenocarcinomas were seen in a fourth low dose male rat and in a second high dose male rat (adenomas or adenocarcinomas, combined: control, 1/50; low dose, 4/50; high dose, 2/50). The historical incidence of renal tubular cell adenomas or adenocarcinomas (combined) in untreated male F344/N rats is 9/1,928 (0.5%), and the highest incidence observed in controls is 3/50. Malignant meningiomas of the brain occurred in 3/50 low dose male rats; none was observed in other groups. The historical incidence of meningiomas in untreated male F344/N rats is 2/1,928 (0.1%). C-Cell adenomas of the thyroid gland in female rats occurred with a positive trend; the incidence in the high dose group was not statistically greater than that in the controls (4/50; 6/50; 11/50). A C-cell carcinoma occurred in another low dose female rat. The incidence of adenomas of the anterior pituitary gland in low dose male rats was marginally greater than that in controls (4/50; 11/50; 8/50). Neither of these marginal increases was considered to be chemically related. Malignant mixed tumors (adenocarcinoma, type C) of the mammary gland occurred in dosed female mice (0/50; 1/50; 5/48). One mammary gland acinar cell carcinoma occurred in a second low dose female mouse. The historical incidence of all malignant mammary gland neoplasms in untreated female B6C3F1 mice is 40/2,040 (2%). Compound-related nonneoplastic lesions of the kidney in mice included nephropathy and dilatation of the renal pelvis for males and females and tubular cysts, suppurative inflammation, and epithelial hyperplasia of the renal pelvis for males. Kidney lesions may have contributed to the low survival of high dose female mice. Mucosal epithelial hyperplasia and submucosal chronic focal inflammation of the urinary bladder were observed at increased incidences in dosed male mice. Suppurative inflammation of the prostate was observed at an increased incidence in high dose male mice. Fighting may have contributed to urogenital lesions in male mice. Suppurative inflammation of the ovary or uterus was observed at an increased incidence in high dose female mice. Hematopoiesis was observed at increased incidences in the spleen and liver of dosed male and high dose female mice and in the adrenal cortex of high dose female mice. Genetic Toxicology: Furosemide was not mutagenic in S. typhimurium strains TA98, TA100, TA1535, or TA1537 when tested with or without exogenous metabolic activation. In the mouse lymphoma assay for trifluorothymidine (Tft) resistance, furosemide produced an equivocal response in the absence of metabolic activation and a positive response in the presence of activation. Furosemide induced sister chromatid exchanges and chromosomal aberrations in CHO cells in both the presence and absence of exogenous metabolic activation. Audit: The data, documents, and pathology materials from the 2-year studies of furosemide have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusion: Under the conditions of these 2-year feed studies, there was equivocal evidence of carcinogenic activity of furosemide for male F344/N rats, as shown by marginal increases in uncommon tubular cell neoplasms of the kidney and meningiomas of the brain. There was no evidence of carcinogenic activity of furosemide for female F344/N rats fed diets containing 350 or 700 ppm furosemide for 2 years. There was no evidence of carcinogenic activity for male B6C3F1 mice fed diets containing 700 or 1,400 ppm furosemide for 2 years. There was some evidence of carcinogenic activity of furosemide for female mice, as shown by an increase in malignant tumors of the mammary gland. Nephropathy was more severe in the kidney of male rats and of male and female mice fed diets containing furosemide than in controls. Synonyms: 5-(aminosulfonyl)-4-chloro-2-[(2-furanylmethyl)amino]benzoic acid; frusemide; fursemide Trade Names: Aisemide; Aluzine; Beronald; Desdemin; Diural; Dryptal; Errolon; Frusemin; Fulsix; Fuluvamide; Furosemide "Mita"; Katlex; Lasilix; Lasix; Lowpstron; Rosemide; Transit; Urosemide


Effects of disinfectant type and dosage on biofilm's activity, viability, microbiome and antibiotic resistome in bench-scale drinking water distribution systems.
Drinking water distribution systems (DWDSs) are important for supplying high-quality water to consumers and disinfectant is widely used to control microbial regrowth in DWDSs. However, the disinfectant's influences on microbial community and antibiotic resistome in DWDS biofilms and the underlying mechanisms driving their dynamics remain elusive. The study investigated the effects of chlorine and chloramine disinfection on the microbiome and antibiotic resistome of biofilms in bench-scale DWDSs using metagenomics assembly. Additionally, the biofilm activity and viability were monitored based on adenosine triphosphate (ATP) and flow cytometer (FCM) staining. The results showed that both chlorine and chloramine disinfectants decreased biofilm ATP, although chloramine at a lower dosage (1 mg/L) could increase it. Chloramine caused a greater decrease in living cells than chlorine. Furthermore, the disinfectants significantly lowered the microbial community diversity and altered microbial community structure. Certain bacterial taxa were enriched, such as Mycobacterium, Sphingomonas, Sphingopyxis, Azospira, and Dechloromonas. Pseudomonas aeruginosa exhibited high resistance towards disinfectants. The disinfectants also decreased the complexity of microbial community networks. Some functional taxa (e.g., Nitrospira, Nitrobacter, Nitrosomonas) were identified as keystones in chloramine-treated DWDS microbial ecological networks. Stochasticity drove biofilm microbial community assembly, and disinfectants increased the contributions of stochastic processes. Chlorine had greater promotion effects on antibiotic resistance genes (ARGs), mobile genetic elements (MGEs) and ARG hosts than chloramine. The disinfectants also selected pathogens, such as Acinetobacter baumannii and Klebsiella pneumonia, and these pathogens also harbored ARGs and MGEs. Overall, this study provides new insights into the effects of disinfectants on biofilm microbiome and antibiotic resistome, highlighting the importance of monitoring and managing disinfection practices in DWDSs.


Spontaneous Droplet Motion on a Periodically Compliant Substrate.
Droplet motion arises in many natural phenomena, ranging from the familiar gravity-driven slip and arrest of raindrops on windows to the directed transport of droplets for water harvesting by plants and animals under dry conditions. Deliberate transportation and manipulation of droplets are also important in many technological applications, including droplet-based microfluidic chemical reactors and for thermal management. Droplet motion usually requires gradients of surface energy or temperature or external vibration to overcome contact angle hysteresis. Here, we report a new phenomenon in which a drying droplet placed on a periodically compliant surface undergoes spontaneous, erratic motion in the absence of surface energy gradients and external stimuli such as vibration. By modeling the droplet as a mass-spring system on a substrate with periodically varying compliance, we show that the stability of equilibrium depends on the size of the droplet. Specifically, if the center of mass of the drop lies at a stable equilibrium point of the system, it will stay there until evaporation reduces its size and this fixed point becomes unstable; with any small perturbation, the droplet then moves to one of its neighboring fixed points.


Analysis and Identification Genetic Effect of SARS-CoV-2 Infections to Alzheimer's Disease Patients by Integrated Bioinformatics.
COVID-19 pandemic is a global crisis which results in millions of deaths and causes long-term neurological sequelae, such as Alzheimer's disease (AD). We aimed to explore the interaction between COVID-19 and AD by integrating bioinformatics to find the biomarkers which lead to AD occurrence and development with COVID-19 and provide early intervention. The differential expressed genes (DEGs) were found by GSE147507 and GSE132903, respectively. The common genes between COVID-19 and AD were identified. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interactions (PPI) network analysis were carried out. Hub genes were found by cytoscape. A multivariate logistic regression model was constructed. NetworkAnalyst was used for the analysis of TF-gene interactions, TF-miRNA coregulatory network, and Protein-chemical Interactions. Forty common DEGs for AD and COVID-19 were found. GO and KEGG analysis indicated that the DEGs were enriched in the calcium signal pathway and other pathways. A PPI network was constructed, and 5 hub genes were identified (ITPR1, ITPR3, ITPKB, RAPGEF3, MFGE8). Four hub genes (ITPR1, ITPR3, ITPKB, RAPGEF3) which were considered as important factors in the development of AD that were affected by COVID-19 were shown by nomogram. Utilizing NetworkAnalyst, the interaction network of 4 hub genes and TF, miRNA, common AD risk genes, and known compounds is displayed, respectively. COVID-19 patients are at high risk of developing AD. Vaccination is required. Four hub genes can be considered as biomarkers for prediction and treatment of AD development caused by COVID-19. Compounds with neuroprotective effects can be used as adjuvant therapy for COVID-19 patients.


Opioid pharmacotherapy for chronic noncancer pain: the american experience.
Chronic noncancer pain is a significant and growing public health challenge in the United States. Lacking effective alternative interventions for effective chronic noncancer pain management, many physicians have turned to opioid pharmacotherapy. Increased opioid prescribing brings not only gains in therapeutic benefit but also a higher incidence of adverse drug events including increased medication misuse and opioid related mortality. Currently the United States must confront the dual problems of widespread undertreated chronic noncancer pain and a prescription opioid abuse crisis. Withholding pain relieving drugs from patients in need is unjustifiable, yet drug diversion, abuse and adverse drug events have become major social as well as medical problems. At the heart of this crisis is the lack of definitive evidence about the risk to benefit ratio of opioid pharmacotherapy for chronic noncancer pain both on an individual case and on a population basis. This article describes the extent and severity of the American chronic noncancer pain problem and the history of opioid pharmacotherapy for chronic noncancer pain in the United States. It then discusses the concept of evidence based practice and reviews current evidence supporting opioid pharmacotherapy for chronic noncancer pain as well as adverse drug events related to opioid pharmacotherapy including misuse and abuse. Finally, it considers the conflict of providing pain relief versus protecting society and reviews steps that governmental agencies, industry and others are taking to contain and ultimately resolve the problems of excessive prescribing and conflicting priorities.


Smoking behavior and delivery of evidence-based care for veterans with spinal cord injuries and disorders.
Little is known about those veterans with spinal cord injuries and disorders (SCI/D) who smoke cigarettes. This study identified the factors associated with smoking in this population, motivations for smoking, and the readiness for smoking cessation. Current practices for the delivery of evidence-based tobacco cessation were also examined. Methods included surveys of veterans with SCI/D, medical record reviews of current smokers, and telephone interviews with SCI/D providers. Six Veterans Health Administration facilities with SCI centers and one SCI clinic. Survey data were analyzed for 1210 veterans, 256 medical records were reviewed, and 15 providers served as key informants. Observational study. Veterans self-reported smoking status, quit attempts, methods and care received, motivation for smoking, and health care conditions. Medical record review and informant interviews examined the tobacco cessation care provided. Whereas 22% of the respondents were current smokers; 51% were past smokers. Current smokers more often reported respiratory illnesses and/or symptoms, alcohol use, pain, and depression than past or never smokers, and approximately half made quit attempts in the past year. Smokers received referral to counseling (57%) and/or prescription for medication/nicotine replacement (23%). Key informants identified difficulty of providing follow-up and patients' unwillingness to consider quitting as barriers. Veterans with SCi/D who smoke also had other health problems. Providers offer counseling and medication, but often have difficulty following patients to arrange/provide support. Identifying other support methods such as telehealth, considering the use of combination cessation therapies, and addressing other health concerns (e.g., depression) may affect tobacco cessation in this population.


Self-interference compressive digital holography with improved axial resolution and signal-to-noise ratio.
Fresnel incoherent correlation holography (FINCH) was proposed to break the barrier of spatial incoherent digital holographic imaging and show the potential of super-resolution imaging preferences. We developed FINCH as a compressive sensing modality and reconstruction procedure as an inverse problem in order to realize 3D tomographic imaging. Improved axial resolution is obtained via compressive reconstruction. Reconstruction guarantees and accuracy of the proposed method are discussed. Compared with the real-valued signal operation, the signal-to-noise ratio of the results is increased when reconstructing from the complex-valued hologram obtained from the FINCH system.


Characterization of the apoptosis-associated tyrosine kinase (AATYK) expressed in the CNS.
We isolated three related cDNA clones from a mouse cerebellar library; the type I cDNA was identical to the gene encoding the apoptosis-associated tyrosine kinase (AATYK), whose expression in myeloid precursor cells is increased during growth arrest or apoptosis. Low levels of AATYK mRNA expression were seen in adult mouse brains but not in embryos. In situ hybridization confirmed the widespread expression of AATYK mRNA in neurons throughout the adult brain. AATYK possessed tyrosine kinase activity and was autophosphorylated when expressed in 293 cells. AATYK mRNA expression was rapidly induced in cultured cerebellar granule cells during apoptosis induced by a low concentration of KCl (5 mM). Levels of endogenous AATYK protein were increased only slightly, but they were accompanied by an increase in molecular weight during apoptosis. Results of the tyrosine phosphatase treatments indicated that the increase in molecular weight was partly caused by tyrosine phosphorylation. The number of apoptotic granule cells overexpressing wild-type AATYK protein was significantly greater than the number of apoptotic granule cells overexpressing a mutant AATYK that lacked tyrosine kinase activity in low concentrations of KCl. These findings suggest that through its tyrosine kinase activity, AATYK is involved in the apoptosis of mature neurons.


The genome sequence of the fork-jawed nomad bee, Nomada ruficornis (Linnaeus, 1758).
We present a genome assembly from an individual male Nomada ruficornis (the fork-jawed nomad bee; Arthropoda; Insecta; Hymenoptera; Apidae). The genome sequence is 273.0 megabases in span. Most of the assembly is scaffolded into 16 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 20.66 kilobases in length.


Clinical and radiographic outcomes of four different treatment strategies in patients with early rheumatoid arthritis (the BeSt study): A randomized, controlled trial.
Several treatment strategies have proven value in the amelioration of rheumatoid arthritis (RA), but the optimal strategy for preventing long-term joint damage and functional decline is unclear. We undertook this study to compare clinical and radiographic outcomes of 4 different treatment strategies, with intense monitoring in all patients. In a multicenter, randomized clinical trial, 508 patients were allocated to 1 of 4 treatment strategies: sequential disease-modifying antirheumatic drug monotherapy (group 1), step-up combination therapy (group 2), initial combination therapy with tapered high-dose prednisone (group 3), and initial combination therapy with the tumor necrosis factor antagonist infliximab (group 4). Treatment adjustments were made every 3 months in an effort to obtain low disease activity (a Disease Activity Score in 44 joints of < or =2.4). Initial combination therapy including either prednisone (group 3) or infliximab (group 4) resulted in earlier functional improvement than did sequential monotherapy (group 1) and step-up combination therapy (group 2), with mean scores at 3 months on the Dutch version of the Health Assessment Questionnaire (D-HAQ) of 1.0 in groups 1 and 2 and 0.6 in groups 3 and 4 (P < 0.001). After 1 year, mean D-HAQ scores were 0.7 in groups 1 and 2 and 0.5 in groups 3 and 4 (P = 0.009). The median increases in total Sharp/Van der Heijde radiographic joint score were 2.0, 2.5, 1.0, and 0.5 in groups 1-4, respectively (P < 0.001). There were no significant differences in the number of adverse events and withdrawals between the groups. In patients with early RA, initial combination therapy including either prednisone or infliximab resulted in earlier functional improvement and less radiographic damage after 1 year than did sequential monotherapy or step-up combination therapy.


Sleep in a comparative context: Investigating how human sleep differs from sleep in other primates.
Primates vary in their sleep durations and, remarkably, humans sleep the least per 24-hr period of the 30 primates that have been studied. Using phylogenetic methods that quantitatively situate human phenotypes within a broader primate comparative context, we investigated the evolution of human sleep architecture, focusing on: total sleep duration, rapid eye movement (REM) sleep duration, non-rapid eye movement (NREM) sleep duration, and proportion of sleep in REM. We used two different Bayesian methods: phylogenetic prediction based on phylogenetic generalized least squares and a multistate Onrstein-Uhlenbeck (OU) evolutionary model of random drift and stabilizing selection. Phylogenetic prediction confirmed that humans sleep less than predicted for a primate of our body mass, predation risk, brain size, foraging needs, sexual selection, and diet. These analyses further revealed that humans pack an unexpectedly higher proportion of REM sleep within a shorter overall sleep duration, and do so by reducing NREM sleep (rather than increasing REM). The OU model generally confirmed these findings, with shifts along the human lineage inferred for TST, NREM, and proportion of REM, but not for REM. We propose that the risks and opportunity costs of sleep are responsible for shorter sleep durations in humans, with risks arising from terrestrial sleep involving threats from predators and conspecifics, and opportunity costs because time spent sleeping could be used for learning, creating material objects, and socializing.


Chronic epididymitis and Grade III varicocele and their associations with semen characteristics in men consulting for couple infertility.
Chronic epididymitis and varicocele are frequently observed genital disorders in men consulting for couple infertility, but their impact on semen characteristics at the time of infertility consultation is still a matter of debate. We investigated 652 male partners of couples who had their first infertility consultation between 1999 and 2015 in Argentina. Men with chronic epididymitis (n = 253), Grade III varicocele (n = 106), and both conditions (n = 125) were compared with a control group (n = 168) composed of men without these disorders or any other recognized causes of male infertility. We showed that men who presented isolated chronic epididymitis were more likely to have high percentages of low sperm motility and abnormal sperm morphology as well as a high number of white blood cells. Men with isolated Grade III varicocele had low sperm production and motility and an increased percentage of abnormal sperm morphology. Finally, men who simultaneously presented chronic epididymitis with Grade III varicocele had a low sperm motility and increased percentage of abnormal sperm morphology as well as a high number of white blood cells. Physical examination of the genital organs may identify common disorders, potentially involved as causal factors of patient's infertility. These disorders are associated with specific seminal profiles that should help in identifying the best treatment from the available therapeutic options, effectiveness, safety, and allowing as much as possible natural conception.


Current and future trends in the anatomical and functional imaging of head and neck paragangliomas.
Head and neck paragangliomas (HNPGLs) account for approximately 3% of all paragangliomas (PGLs). Most often, HNPGLs are benign, nonsecreting, and slowly progressing. The initial physical examination and biochemical diagnosis usually adds very little to the proper diagnosis of these tumors, and, therefore, radiologists and nuclear medicine physicians play a pivotal role in providing the initial diagnosis, the locoregional staging, and the plan for detecting potential multicentric or metastatic lesions. Based on several current studies, the most accurate use of HNPGL-specific initial and subsequent imaging modalities must be guided by the knowledge of genetics and the specifically measured biochemical profile of these tumors for the proper management of these patients. Thus, this short review article presents the application of the most up-to-date anatomical and functional imaging approaches to HNPGLs tightly linked to the clinical management of these patients. Based on the most recent studies, 18F-FDOPA PET/CT has been shown to be a useful addition to anatomical imaging in the preoperative localization and molecular assessment of HNPGLs. It is estimated that the frequency of metabolically active PGLs on 18F-FDOPA PET/CT in this region is higher than 90%. For patients with hereditary PGL syndromes, (18)F-FDG-PET/CT should be reserved. Imaging of somatostatin receptors using Octreoscan or 68Ga-labeled somatostatin analogues plays an important role for selecting patients for targeted radiation therapy. This review also concludes that it is expected that in the near future, these patients will indeed benefit from new diagnostic approaches based on the identification of new targets by molecular profiling studies that will result in the development of novel PGL-specific radiopharmaceuticals.


Increase in reactive nitrogen species production in chronic obstructive pulmonary disease airways.
Peroxynitrite, nitrogen dioxide, and other reactive nitrogen species (RNS) that are formed in the reaction of nitric oxide (NO) with superoxide anion, and in peroxidase-dependent mechanisms, have a potent inflammatory action. These molecules may therefore increase in number and have a role in inflammatory airway diseases. In the present study, we quantified RNS using immunostaining of nitrotyrosine and inducible NO synthase (iNOS) in airway inflammatory cells obtained by the induced sputum technique, and also quantified the exhaled NO concentration in subjects with chronic obstructive pulmonary disease (COPD), subjects with asthma, and healthy subjects (HS). Immunoreactivity for iNOS observed in the airway inflammatory cells was significantly and similarly higher in subjects with COPD and asthma than in HS, although exhaled NO levels were increased only in subjects with asthma. Inflammatory cells showed obvious nitrotyrosine immunoreactivity in subjects with COPD and to a lesser extent in those with asthma, but not in HS. There was a significant negative correlation between the percent predicted values of FEV(1) and the amount of nitrotyrosine formation in subjects with COPD, but not in those with asthma and HS. These results suggest that: (1) RNS may be involved in the pathobiology of the airway inflammatory and obstructive process in COPD; and (2) NO produced in the airways, presumably via iNOS, is consumed by its reaction with superoxide anion and/or peroxidase-dependent mechanisms.


Cognitive deficits in the Steele-Richardson-Olszewski syndrome (progressive supranuclear palsy).
Clinical examination of 33 consecutive newly diagnosed cases of Steele-Richardson-Olszewski syndrome revealed evidence of cognitive impairment in 20. Eighteen of the 27 right handed patients who underwent neuropsychological assessment had intellectual impairment (mild in nine and marked in nine patients). The pattern of abnormalities was similar to but more severe than those previously reported in Parkinson's disease, with particular difficulties in carrying out tests which are believed to be sensitive to frontal lobe dysfunction.


Intravenous immunoglobulin reduces anti-HLA alloreactivity and shortens waiting time to cardiac transplantation in highly sensitized left ventricular assist device recipients.
Recipients of left ventricular assist devices (LVADs) develop prominent B-cell hyperreactivity. We investigated the influence of anti-HLA antibodies on waiting time to cardiac transplantation in LVAD recipients and compared the effects of 2 immunomodulatory regimens on anti-HLA serum reactivity. Fifty-five previously nonsensitized LVAD recipients of a TCI device implanted between 1990 and 1996 were studied. Patients with anti-HLA antibodies received monthly courses of either intravenous immunoglobulin (IVIg) or plasmapheresis, in conjunction with cyclophosphamide. The effects of these regimens on anti-HLA alloreactivity and waiting time to transplantation were then determined by Kaplan-Meier log-rank statistics, nonparametric Wilcoxon rank-sum test, and Student's t test. Prolongation in transplant waiting time was related to serum IgG anti-HLA class I alloreactivity. Infusion of IVIg (2 g/kg) caused a mean reduction of 33% in anti-HLA class I alloreactivity within 1 week. Waiting time to transplantation was significantly reduced by IVIg therapy and subsequently approximated that in nonsensitized patients. Side effects of IVIg (2 g/kg) were minimal and related primarily to immune complex disease. Although plasmapheresis caused a similar reduction in alloreactivity to IVIg, this effect was achieved after longer treatment. Moreover, plasmapheresis was associated with an unacceptably high frequency of infectious complications. In patients resistant to low-dose (2 g/kg) IVIg therapy, high-dose (3 g/kg) IVIg was effective in reducing alloreactivity but was associated with a high incidence of reversible renal insufficiency. These results indicate that IVIg is an effective and safe modality for sensitized recipients awaiting cardiac transplantation, reducing serum anti-HLA alloreactivity and shortening the duration to transplantation. The therapeutic and safety profile of IVIg would appear to be superior to plasmapheresis.


Accidental intra-arterial injection of diclofenac -case report.
Diclofenac sodium is a very common drug used in medical practice for analgesia and is freely available over the counter without any medical prescription. The drug is also the most commonly used or misused by quacks working all over India. Many case reports have been published on upper limb catastrophe of unintentional intra-arterial injection of barbiturates, thiopental sodium, narcotics, and tranquilizers but only two cases of inadvertent intra-arterial injection of diclofenac have been reported till date. Potential serious complication of inadvertent intra-arterial injection of diclofenac sodium leading to gangrene has been recently reported in medical literature. It is a medical emergency and literature has shown that timely diagnosis with early intervention can salvage the limb. We hereby report a case of accidental intra-arterial injection of diclofenac sodium in the ulnar artery by a quack for pain abdomen leading to gangrene of the distal part of affected little, ring and part of middle finger of right hand.


Development of an SPRi Test for the Quantitative Detection of Cadherin 12 in Human Plasma and Peritoneal Fluid.
A new method for the determination of cadherin 12 (CDH12)-an adhesive protein that has a significant impact on the development, growth, and movement of cancer cells-was developed and validated. The method is based on a biosensor using surface plasmon resonance imaging (SPRi) detection. A quartz crystal microbalance was used to analyze the characteristics of the formation of successive layers of the biosensor, from the linker monolayer to the final capture of CDH12 from solution. The association equilibrium constant (KA = 1.66 × 1011 dm3 mol-1) and the dissociation equilibrium constant (KD = 7.52 × 10-12 mol dm-3) of the anti-CDH12 antibody-CDH12 protein complex were determined. The determined analytical parameters, namely the values determining the accuracy, precision, and repeatability of the method, do not exceed the permissible 20% deviations specified by the aforementioned institutions. The proposed method is also selective with respect to possible potential interferents, occurring in up to 100-fold excess concentration relative to the CDH12 concentration. The determined Limit of Quantification (LOQ = 4.92 pg mL-1) indicates the possibility of performing quantitative analysis in human plasma or peritoneal fluid without the need to concentrate the samples; however, particular attention should be paid to their storage conditions, as the analyte does not exhibit high stability. The Passing-Bablok regression model revealed good agreement between the reference method and the SPRi biosensor, with ρSpearman values of 0.961 and 0.925.


Humor therapy: relieving chronic pain and enhancing happiness for older adults.
The present study examined the effectiveness of a humor therapy program in relieving chronic pain, enhancing happiness and life satisfaction, and reducing loneliness among older persons with chronic pain. It was a quasiexperimental pretest-posttest controlled design. Older persons in a nursing home were invited to join an 8-week humor therapy program (experimental group), while those in another nursing home were treated as a control group and were not offered the program. There were 36 older people in the experimental group and 34 in the control group. Upon completion of the humor therapy program, there were significant decreases in pain and perception of loneliness, and significant increases in happiness and life satisfaction for the experimental group, but not for the control group. The use of humor therapy appears to be an effective nonpharmacological intervention. Nurses and other healthcare professionals could incorporate humor in caring for their patients.


Are different facets of plant diversity well protected against climate and land cover changes? A test study in the French Alps.
Climate and land cover changes are important drivers of the plant species distributions and diversity patterns in mountainous regions. Although the need for a multifaceted view of diversity based on taxonomic, functional and phylogenetic dimensions is now commonly recognized, there are no complete risk assessments concerning their expected changes. In this paper, we used a range of species distribution models in an ensemble-forecasting framework together with regional climate and land cover projections by 2080 to analyze the potential threat for more than 2,500 plant species at high resolution (2.5 km × 2.5 km) in the French Alps. We also decomposed taxonomic, functional and phylogenetic diversity facets into α and β components and analyzed their expected changes by 2080. Overall, plant species threats from climate and land cover changes in the French Alps were expected to vary depending on the species' preferred altitudinal vegetation zone, rarity, and conservation status. Indeed, rare species and species of conservation concern were the ones projected to experience less severe change, and also the ones being the most efficiently preserved by the current network of protected areas. Conversely, the three facets of plant diversity were also projected to experience drastic spatial re-shuffling by 2080. In general, the mean α-diversity of the three facets was projected to increase to the detriment of regional β-diversity, although the latter was projected to remain high at the montane-alpine transition zones. Our results show that, due to a high-altitude distribution, the current protection network is efficient for rare species, and species predicted to migrate upward. Although our modeling framework may not capture all possible mechanisms of species range shifts, our work illustrates that a comprehensive risk assessment on an entire floristic region combined with functional and phylogenetic information can help delimitate future scenarios of biodiversity and better design its protection.


Regulation of arginase II by interferon regulatory factor 3 and the involvement of polyamines in the antiviral response.
The innate antiviral response requires the induction of genes and proteins with activities that limit virus replication. Among these, the well-characterized interferon beta (IFNB) gene is regulated through the cooperation of AP-1, NF-kappaB and interferon regulatory factor 3 (IRF-3) transcription factors. Using a constitutively active form of IRF-3, IRF-3 5D, we showed previously that IRF-3 also regulates an IFN-independent antiviral response through the direct induction of IFN-stimulated genes. In this study, we report that the arginase II gene (ArgII) as well as ArgII protein concentrations and enzymatic activity are induced in IRF-3 5D-expressing and Sendai virus-infected Jurkat cells in an IFN-independent manner. ArgII is a critical enzyme in the polyamine-biosynthetic pathway. Of the natural polyamines, spermine possesses antiviral activity and mediates apoptosis at physiological concentrations. Measurement of intracellular polyamine content revealed that expression of IRF-3 5D induces polyamine production, but that Sendai virus and vesicular stomatitis virus infections do not. These results show for the first time that the ArgII gene is an early IRF-3-regulated gene, which participates in the IFN-independent antiviral response through polyamine production and induction of apoptosis.


Effects of a community-based progressive resistance training program on muscle performance and physical function in adults with Down syndrome: a randomized controlled trial.
To determine whether progressive resistance training improves muscle strength, muscle endurance, and physical function in adults with Down syndrome. Single-blind randomized controlled trial. General community. Adults (N=20) with Down syndrome (13 men, 7 women; mean age, 26.8+/-7.8 y) were randomly assigned through a concealed allocation block randomized method to either an intervention group (n=9) or a control group (n=11). The intervention was a supervised, group progressive resistance training program, consisting of 6 exercises using weight machines performed twice a week for 10 weeks. Participants completed 2 to 3 sets of between 10 to 12 repetitions of each exercise until they reached fatigue. The control group continued with their usual activities. The outcomes measured by blinded assessors were muscle strength (1-repetition maximum [1-RM]), muscle endurance (number of repetitions at 50% of 1-RM) for chest press and leg press, timed stairs test, and the grocery shelving task. The intervention group showed significant improvement in upper-limb muscle endurance compared with the control group (mean difference in the number of repetitions of the chest press at 50% of 1-RM was 16.7, 95% confidence interval, [CI] 7.1-26.2); and a trend toward an improvement in upper-limb muscle strength (mean difference in chest press 1-RM, 8.6 kg; 95% CI, -1.3 to 18.5 kg) and in upper-limb function (mean difference in grocery shelving task, -20.3s; 95% CI, -45.7 to 5.2s). There were no significant differences between the groups for lower-limb muscle performance or physical function measures. No major adverse events for the intervention were noted. Progressive resistance training is a safe and feasible fitness option that can improve upper-limb muscle endurance in adults with Down syndrome (ACTR identifier ACTRN 012606000515594.).


Intramedullary Neurocysticercosis Mimicking Cord Tumor.
Isolated spinal intramedullary involvement by neurocysticercosis is extremely rare. We report a case of a Hispanic female with right-sided weakness, magnetic resonance imaging showing cervical intramedullary lesion. Surgery was performed due to the progressive nature of symptoms. The cervical cord lesion was completely removed; pathology was consistent with degenerated cysticercosis. Progressive clinical improvement with physiotherapy was achieved. Although rare, especially in the absence of intracranial lesions, the diagnosis should be considered in appropriate patient population as it usually presents a peripherally enhancing cystic lesion.


Antimicrobial Mechanisms and Preparation of Antibiotic-impregnated Cement-coated Locking Plates in the Treatment of Infected Non-unions.
Antibiotic-impregnated cement-coated plates (ACPs) have been used successfully for temporary internal fixation between stages in the two-stage treatment of infected non-unions. We describe our approach of using an ACP in the staged treatment of a methicillin-resistant Staphylococcus aureus (MRSA)-infected distal femoral non-union below a total hip prosthesis. In addition, we present the results of an in vitro experiment to provide an in-depth insight into the capacity of ACPs in (i) treating residual biofilm and (ii) preventing bacterial recolonisation. In the first stage, we used a titanium LISS plate coated with hand-mixed PALACOS with vancomycin (PAL-V) for temporary internal fixation combined with commercially prepared COPAL with gentamicin and vancomycin (COP-GV) to fill the segmental defect. In the second stage, the non-union was treated with double-plate fixation and bone grafting.A Kirby-Bauer agar disc diffusion assay was performed to determine the antimicrobial activity of both ACPs and a drug-release assay to measure antibiotic release over time. A biofilm killing assay was also carried out to determine if the antibiotic released was able to reduce or eradicate biofilm of the patient's MRSA strain. At one-year follow-up, there was complete bone-bridging across the previous non-union. The patient was pain-free and ambulatory without need for further surgery. Both ACPs with COP-GV and PAL-V exerted an antimicrobial effect against the MRSA strain with peak concentrations of antibiotic released within the first 24 hours. Concentrations released from COP-GV in the first 24 hours in vitro caused a 7.7-fold log reduction of colony-forming units (CFU) in the biofilm. At day 50, both COP-GV and PAL-V still released concentrations of antibiotic above the respective minimal inhibitory concentrations (MIC), likely contributing to the positive clinical outcome. The use of an ACP provides stability and infection control in the clinical scenario of an infected non-union. This is confirmed in vitro where the release of antibiotics from ACPs is characterised by an early burst followed by a prolonged sustained release above the MIC until 50 days. The burst release from COP-GV reduces CFU in the biofilm and prevents early recolonisation through synergistic activity of the released vancomycin and gentamicin. An antibiotic-impregnated cement-coated plate is a useful addition to the surgeon's armamentarium to provide temporary internal fixation without the disadvantages of external fixation and contribute to infection control in an infected non-union. Wagner RK, Guarch-Pérez C, van Dam AP, et al. Antimicrobial Mechanisms and Preparation of Antibiotic-impregnated Cement-coated Locking Plates in the Treatment of Infected Non-unions. Strategies Trauma Limb Reconstr 2023;18(2):73-81.


Surface characterization and platelet adhesion studies for the mixed self-assembled monolayers with amine and carboxylic acid terminated functionalities.
The mixed self-assembled monolayers (SAM) prepared from long chain alkanethiols, HS(CH(2))(11)NH(2) and HS(CH(2))(10)COOH, on gold are employed as the model surface for investigating the interactions between the biological environment and synthetic surface. A distinctive SAM preparation scheme was utilized in this investigation. The triethylamine was added to the alkanethiol solution during SAM formation and then followed by additional rinsing of SAM with 10% CH(3)COOH or 1% HCl ethanolic solution. The contact angle values of NH(2) + COOH mixed SAMs were between those of the pure SAMs, except that it was prepared with solution mole fraction of amine-terminated alkanethiol at 0.2. X-ray photoelectron spectroscopy (XPS) analysis has indicated that these two distinctive SAM preparation procedures had both resulted in a reduction in oxidized sulfur species on pure --NH(2) terminated SAM. However, the procedure utilizing 1% HCl ethanolic washing solution was more effective in reducing the unbound thiol fraction and to form a pure --NH(2) SAM with better quality. XPS analysis has also revealed that the surface of NH(2) + COOH mixed SAMs was "amine-rich". In vitro platelet adhesion assay has shown that the amount of adherent platelets on pure positive charged --NH(2) terminated SAM is less than that on anionic --COOH terminated counterpart in both acidic ethanolic washing schemes. Moreover, the lowest platelet adhesion density was noted on the mixed SAM surfaces with surface amine mole fraction at 0.51 and 0.57. This finding suggests that the surface charge with near neutrality might be of importance in reducing platelet adhesion and activation on artificial biomaterial.


TRIM27 acts as an oncogene and regulates cell proliferation and metastasis in non-small cell lung cancer through SIX3-β-catenin signaling.
The Wnt/β-catenin pathway plays vital roles in diverse biological processes, including cell differentiation, proliferation, migration, and insulin sensitivity. A recent study reported that the DNA-binding transcriptional factor SIX3 is essential during embryonic development in vertebrates and capable of downregulating target genes of the Wnt/β-catenin pathway in lung cancer, indicating negative regulation of Wnt/β-catenin activation. However, regulation of the SIX3-Wnt/β-catenin pathway axis remains unknown. We measured the expression of TRIM27 and SIX3 as well as investigated whether there was a correlation between them in lung cancer tissue samples. Herein, we found that the E3 ubiquitin ligase, TRIM27, ubiquitinates, and degrades SIX3. TRIM27 induces non-small cell lung cancer (NSCLC) cell proliferation and metastasis, and the expression of β-catenin, S100P, TGFB3, and MMP-9 were significantly inhibited by SIX3. Furthermore, XAV939 is a selective β-catenin-mediated transcription inhibitor that inhibited TRIM27- and SIX3-mediated NSCLC cell proliferation, migration, and invasion. Clinically, lung tissue samples of cancer patients showed increased TRIM27 expression and decreased SIX3 expression. Taken together, these data demonstrate that TRIM27 acts as an oncogene regulating cell proliferation and metastasis in NSCLC through SIX3-β-catenin signaling.


Molecular mechanisms of hormone-mediated Müllerian duct regression: involvement of beta-catenin.
Regression of the Müllerian duct in the male embryo is one unequivocal effect of anti-Müllerian hormone, a glycoprotein secreted by the Sertoli cells of the testis. This hormone induces ductal epithelial regression through a paracrine mechanism originating in periductal mesenchyme. To probe the mechanisms of action of anti-Müllerian hormone, we have studied the sequence of cellular and molecular events involved in duct regression. Studies were performed in male rat embryos and in transgenic mice overexpressing or lacking anti-Müllerian hormone, both in vivo and in vitro. Anti-Müllerian hormone causes regression of the cranial part of the Müllerian duct whereas it continues to grow caudally. Our work shows that this pattern of regression is correlated with a cranial to caudal gradient of anti-Müllerian hormone receptor protein, followed by a wave of apoptosis spreading along the Müllerian duct as its progresses caudally. Apoptosis is also induced by AMH in female Müllerian duct in vitro. Furthermore, apoptotic indexes are increased in Müllerian epithelium of transgenic mice of both sexes overexpressing the human anti-Müllerian hormone gene, exhibiting a positive correlation with serum hormone concentration. Inversely, apoptosis is reduced in male anti-Müllerian hormone-deficient mice. We also show that apoptosis is a decisive but not sufficient process, and that epitheliomesenchymal transformation is an important event of Müllerian regression. The most striking result of this study is that anti-Müllerian hormone action in peri-Müllerian mesenchyme leads in vivo and in vitro to an accumulation of cytoplasmic beta-catenin. The co-localization of beta-catenin with lymphoid enhancer factor 1 in the nucleus of peri-Müllerian mesenchymal cells, demonstrated in primary culture, suggests that overexpressed beta-catenin in association with lymphoid enhancer factor 1 may alter transcription of target genes and may lead to changes in mesenchymal gene expression and cell fate during Müllerian duct regression. To our knowledge, this is the first report that beta-catenin, known for its role in Wnt signaling, may mediate anti-Müllerian hormone action.


Adsorbate-induced structural evolution changes the mechanism of CO oxidation on a Rh/Fe3O4(001) model catalyst.
The structure of a catalyst often changes in reactive environments, and following the structural evolution is crucial for the identification of the catalyst's active phase and reaction mechanism. Here we present an atomic-scale study of CO oxidation on a model Rh/Fe3O4(001) "single-atom" catalyst, which has a very different evolution depending on which of the two reactants, O2 or CO, is adsorbed first. Using temperature-programmed desorption (TPD) combined with scanning tunneling microscopy (STM) and X-ray photoelectron spectroscopy (XPS), we show that O2 destabilizes Rh atoms, leading to the formation of RhxOy clusters; these catalyze CO oxidation via a Langmuir-Hinshelwood mechanism at temperatures as low as 200 K. If CO adsorbs first, the system is poisoned for direct interaction with O2, and CO oxidation is dominated by a Mars-van-Krevelen pathway at 480 K.


Anticancer Study of a Novel Pan-HDAC Inhibitor MPT0G236 in Colorectal Cancer Cells.
Colorectal cancer (CRC) is one of the most commonly diagnosed malignancies and a leading cause of cancer worldwide. Histone deacetylases (HDACs), which regulate cell proliferation and survival, are associated with the development and progression of cancer. Moreover, HDAC inhibitors are promising therapeutic targets, with five HDAC inhibitors approved for cancer treatment to date. However, their safety profile necessitates the exploration of well-tolerated HDAC inhibitors that can be used in cancer therapeutic strategies. In this study, the pan-HDAC inhibitor MPT0G236 reduced the viability and inhibited the proliferation of human colorectal cancer cells, and normal human umbilical vein endothelial cells (HUVECs) showed reduced sensitivity. These findings indicated that MPT0G236 specifically targeted malignant tumor cells. Notably, MPT0G236 significantly inhibited the activities of HDAC1, HDAC2, and HDAC3, Class I HDACs, as well as HDAC6, a Class IIb HDAC, at low nanomolar concentrations. Additionally, it promoted the accumulation of acetyl-α-tubulin and acetyl-histone H3 in HCT-116 and HT-29 cells in a concentration-dependent manner. Furthermore, MPT0G236 treatment induced G2/M cell cycle arrest in CRC cells by initially regulating the levels of cell-cycle-related proteins, such as p-MPM2; specifically reducing p-cdc2 (Y15), cyclin B1, and cdc25C levels; and subsequently inducing apoptosis through the caspase-dependent pathways and PARP activation. Our findings demonstrate that MPT0G236 exhibits significant anticancer activity in human colorectal cancer cells.


Rationale and efficacy of conservative therapy for gastroesophageal reflux disease.
Major recent advances in the diagnosis and understanding of gastroesophageal reflux disease have afforded the practicing clinician a number of therapeutic options to treat the increasingly recognized affected patient population. With highly effective acid-suppressive therapies available, simple life-style modifications, such as selective food and medicine avoidance, weight loss, smoking cessation, elevation of the head of the bed, and the judicious use of antacids and alginate, have been relegated to a minor, if any, role in the management of these patients. The validity of these recommendations, however, remains consistent with our current understanding of the pathogenesis of gastroesophageal reflux disease. Although few well-designed placebo-controlled trials have been conducted, a review of the medical literature indicates an appreciable efficacy of these interventions, which are founded on well-studied physiologic determinants of gastroesophageal reflux. Most patients with gastroesophageal reflux disease can be managed by reassurance and simple life-style adjustments alone. The therapy for those with chronic or relapsing disease should always include the adjunctive reinforcement of these simple, efficacious, and cost-effective measures.


Extended interval BNT162b2 vaccination enhances peak antibody generation.
The BNT162b2 vaccine is highly effective against COVID-19 infection and was delivered with a 3-week time interval in registration studies1. However, many countries extended this interval to accelerate population coverage with a single vaccine. It is not known how immune responses are influenced by delaying the second dose. We provide the assessment of immune responses in the first 14 weeks after standard or extended-interval BNT162b2 vaccination and show that delaying the second dose strongly boosts the peak antibody response by 3.5-fold in older people. This enhanced antibody response may offer a longer period of clinical protection and delay the need for booster vaccination. In contrast, peak cellular-specific responses were the strongest in those vaccinated on a standard 3-week vaccine interval. As such, the timing of the second dose has a marked influence on the kinetics and magnitude of the adaptive immune response after mRNA vaccination in older people.


Caudal analgesia in children. Influence on ventilatory efficiency during halothane anaesthesia.
The influence of caudal analgesia on pulmonary ventilation and gas exchange was studied in 26 children during halothane anaesthesia with spontaneous breathing. Two groups of children were studied with 13 patients in each group. One group received caudal analgesia. The other group had no caudal blocks. All children were subjected to lower abdominal and genital surgical procedures. Minute ventilation and respiratory rates were significantly lower in the caudal group than in the non-caudal group. Wastea minute ventilation and VD/VT ratios were increased in the non-caudal group. The end tidal carbon dioxide concentration was unchanged in both groups. The lower minute ventilation in the caudal group eliminated the same amount or even greater amounts of CO2 per minute indicating an improved gas distribution at slow respiratory rates. The improved ventilation efficiency and the excellent immediate postoperative pain relief achieved by caudal analgesia justifies its frequent use for these operative surgical procedures.


Modifications at K31 on the lateral surface of histone H4 contribute to genome structure and expression in apicomplexan parasites.
An unusual genome architecture characterizes the two related human parasitic pathogens Plasmodium falciparum and Toxoplasma gondii. A major fraction of the bulk parasite genome is packaged as transcriptionally permissive euchromatin with few loci embedded in silenced heterochromatin. Primary chromatin shapers include histone modifications at the nucleosome lateral surface close to the DNA but their mode of action remains unclear. We now identify versatile modifications at Lys31 within the globular domain of histone H4 that crucially determine genome organization and expression in Apicomplexa parasites. H4K31 acetylation at the promoter correlates with, and perhaps directly regulates, gene expression in both parasites. By contrast, monomethylated H4K31 is enriched in the core body of T. gondii active genes but inversely correlates with transcription, whereas it is unexpectedly enriched at transcriptionally inactive pericentromeric heterochromatin in P. falciparum, a region devoid of the characteristic H3K9me3 histone mark and its downstream effector HP1.


Complex study on compression of ECG signals using novel single-cycle fractal-based algorithm and SPIHT.
Compression of ECG signal is essential especially in the area of signal transmission in telemedicine. There exist many compression algorithms which are described in various details, tested on various datasets and their performance is expressed by different ways. There is a lack of standardization in this area. This study points out these drawbacks and presents new compression algorithm which is properly described, tested and objectively compared with other authors. This study serves as an example how the standardization should look like. Single-cycle fractal-based (SCyF) compression algorithm is introduced and tested on 4 different databases-CSE database, MIT-BIH arrhythmia database, High-frequency signal and Brno University of Technology ECG quality database (BUT QDB). SCyF algorithm is always compared with well-known algorithm based on wavelet transform and set partitioning in hierarchical trees in terms of efficiency (2 methods) and quality/distortion of the signal after compression (12 methods). Detail analysis of the results is provided. The results of SCyF compression algorithm reach up to avL = 0.4460 bps and PRDN = 2.8236%.


Hydropower's Biogenic Carbon Footprint.
Global warming is accelerating and the world urgently needs a shift to clean and renewable energy. Hydropower is currently the largest renewable source of electricity, but its contribution to climate change mitigation is not yet fully understood. Hydroelectric reservoirs are a source of biogenic greenhouse gases and in individual cases can reach the same emission rates as thermal power plants. Little is known about the severity of their emissions at the global scale. Here we show that the carbon footprint of hydropower is far higher than previously assumed, with a global average of 173 kg CO2 and 2.95 kg CH4 emitted per MWh of electricity produced. This results in a combined average carbon footprint of 273 kg CO2e/MWh when using the global warming potential over a time horizon of 100 years (GWP100). Nonetheless, this is still below that of fossil energy sources without the use of carbon capture and sequestration technologies. We identified the dams most promising for capturing methane for use as alternative energy source. The spread among the ~1500 hydropower plants analysed in this study is large and highlights the importance of case-by-case examinations.


Method variables affecting the bioaccessibility of arsenic in soil.
Arsenic bioaccessibility tests are now being commonly used in risk assessment. However, concerns remain about the reliability of such tests because the bioaccessibility of arsenic from soil may be susceptible to soil composition (including iron concentration), as well as method considerations such as varying liquid-to-solid ratios and the chosen buffer system. In this study, arsenic-contaminated tailings and soils were tested to compare two bioaccessibility methods: one that uses glycine as a buffer, and a second that is more physiologically based. With the glycine-buffered method, arsenic and iron bioaccessibility increased in the presence of a higher buffer concentration at higher liquid-to-solid ratios, whereas the results of physiologically-based tests were unaffected by variations in these parameters. In the glycine-buffered system, interactions between iron and glycine may influence the concentration of arsenic in solution, which may not be consistent with human gastrointestinal conditions. The choice of a physiologically-based method may be more appropriate to achieve representative arsenic bioaccessibility values toward estimating risks to human health.


Temporal distribution of blood donations in three Brazilian blood centers and its repercussion on the blood supply.
Seasonal distribution of blood donation hinders efforts to provide a safe and adequate blood supply leading to chronic and persistent shortages. This study examined whether holidays, geographical area and donation type (community versus replacement) has any impact on the fluctuation of donations. The numbers of blood donations from 2007 through 2010 in three Brazilian Retrovirus Epidemiological Donor Study II (REDS-II) participating centers were analyzed according to the week of donation. The weeks were classified as holiday or non-holiday. To compare donations performed during holiday versus non-holiday weeks, tabulations and descriptive statistics for weekly donations by blood center were examined and time series analysis was conducted. The average weekly number of donations varied according to the blood center and type of week. The average number of donations decreased significantly during Carnival and Christmas and increased during the Brazilian National Donor Week. The fluctuation was more pronounced in Recife and Belo Horizonte when compared to São Paulo and higher among community donors. National bank holidays affect the blood supply by reducing available blood donations. Blood banks should take into account these oscillations in order to plan local campaigns, aiming at maintaining the blood supply at acceptable levels.


Iron fortification: country level experiences and lessons learned.
Iron fortification has been used to enhance iron intake in many developed countries for more than 50 years, but only in the last decade has this strategy been applied on a large scale to other parts of the world. Iron fortification of rice is being instituted in the Philippines. Initially, the rice will be produced in government-controlled rice mills and sold at low cost mainly to low income families. Efforts to improve the technology (using coating or extrusion techniques) are currently underway to reduce cost and minimize losses during storage and washing. Effectiveness and feasibility studies are required to test the new technologies and processing/distribution systems. In Venezuela in 1993, the government instituted a mandatory program of iron fortification to enrich precooked corn flour followed by the voluntary fortification of wheat flour. Surveys in school children subsequently showed a sharp drop in iron deficiency. Fortification of fish sauce in Vietnam has shown promising initial results in reducing anemia among anemic, nonpregnant female factory workers. Iron-fortified soy sauce has been shown to reduce anemia in initial studies in children in China, and a large-scale efficacy trial is now underway. These examples indicate that iron fortification of staple foods and condiments holds great promise for the prevention of iron deficiency.


Behavior and neural basis of near-optimal visual search.
The ability to search efficiently for a target in a cluttered environment is one of the most remarkable functions of the nervous system. This task is difficult under natural circumstances, as the reliability of sensory information can vary greatly across space and time and is typically a priori unknown to the observer. In contrast, visual-search experiments commonly use stimuli of equal and known reliability. In a target detection task, we randomly assigned high or low reliability to each item on a trial-by-trial basis. An optimal observer would weight the observations by their trial-to-trial reliability and combine them using a specific nonlinear integration rule. We found that humans were near-optimal, regardless of whether distractors were homogeneous or heterogeneous and whether reliability was manipulated through contrast or shape. We present a neural-network implementation of near-optimal visual search based on probabilistic population coding. The network matched human performance.


The politics of lesbian specificity.
The notion of lesbian specificity must be understood in the context of heteronormative society. Heterosexual norms affect negotiations of power and sexual desire in lesbian relationships. But this investigation into woman-to-woman sexual practices also views the possibility of lesbian specificity as potentially subversive, in the sense that it poses fundamental questions about conventional discourses on sexuality and eroticism.


Surgical creation of a Cupid's bow using W-plasty in patients after cleft lip surgery.
The three-dimensional contour of the Cupid's bow is extremely important from a cosmetic standpoint, and many patients with bilateral or unilateral cleft lip require revision or reconstruction of the Cupid's bow.A number of surgical techniques have been reported for creating the Cupid's bow, among which the methods of Gillies and Onizuka are the most widely known. In the methods of Gillies and Onizuka, the dog-ear can result in excessive volume in the vermilion notch, raising the trough in the Cupid's bow and leading to regression during the post-surgical period. We have improved upon these techniques using the W-plasty and have obtained satisfactory results that preserve the white skin roll. In our method, the left and right prominence of the Cupid's bow is created through an incision of the white lip, while the central trough is created through a curvilinear incision of the vermilion mucosa. At the two sites of the white lip tissue and the curvilinear incision of the central vermilion tissue, the white skin roll is preserved as much as possible. In patients where the Gillies method had been used, the Cupid's bow tended to be flattened gradually after surgery. We report on the details of our method and the case reports using the technique.


The safety of synthetic zeolites used in detergents.
Synthetic zeolites are replacing phosphates as builders in laundry detergents; workers and consumers may, therefore, increasingly be exposed to these materials and it is important to assess their safety. This article puts mechanistic, toxicological and exposure data into context for a safety assessment. Zeolites are hygroscopic compounds with ion-exchanging properties. They may partially decompose under acidic conditions such as in the stomach releasing sodium ions, silicic acid and aluminum salts. The intact molecule is not bioavailable after oral intake or exposure through the dermal and inhalational routes. Under current conditions of manufacture and use, no systemic toxicity is to be expected from neither the intact molecule nor the degradation products; a significant effect on the bioavailability of other compounds is not likely. Zeolites may cause local irritation. It is, therefore, important to minimise occupational exposure. The co-operation of detergent manufacturers with the manufacturers of washing machines is necessary to find the right balance between environmental aspects such as energy and water savings and the occurrence of detergent residues on textiles due to insufficient rinsing.


Genetic variants in the leukemia-associated Rho guanine nucleotide exchange factor (ARHGEF12) gene are not associated with T2DM and related parameters in Caucasians (KORA study).
The aim of our study was to determine the variant pattern of the leukemia-associated Rho guanine nucleotide exchange factor (LARG, or ARHGEF12) gene and investigate whether LARG variants are associated with diabetes mellitus type 2 (T2DM), the metabolic syndrome (MetS), or related parameters such as insulin sensitivity in German Caucasians. We analyzed single nucleotide polymorphisms (SNPs) in the LARG gene in the 55-74-year-old individuals of the population-based German Caucasian Cooperative Health Research in the region of Augsberg (KORA) survey 4 (S4). Sequencing of Tyr1306Cys, which was of functional relevance in Pima Indians, in 48 randomly selected individuals and genotyping of 11 additional LARG SNPs in 1653 subjects were performed. Four linkage disequilibrium (LD) blocks (r(2)> or =0.8) were established and each block was statistically analyzed for association with metabolic traits. The association with T2DM and the MetS was analyzed by logistic regression in 1462 subjects, and HOMA-IR (homeostasis model assessment of insulin resistance) as a measure of insulin sensitivity was analyzed by the Kruskal-Wallis test in 1346 fasting subjects. The polymorphism Tyr1306Cys, which was significantly associated with insulin sensitivity in Pima Indians, was not found in the KORA S4 population. Statistical analysis yielded no significant associations (P>0.05) between the analyzed LARG variants and T2DM, the MetS, or related parameters such as insulin sensitivity. Caucasian individuals and Pima Indians differ in their genetic variance pattern in the LARG gene region. There is no evidence in the Caucasian KORA study that variants of the LARG gene confer susceptibility for T2DM, insulin sensitivity, or the MetS.


Validation of preoperative variables and stratification of patients to help predict benefit of cytoreductive nephrectomy in the targeted therapy ERA.
To further elucidate which patients with metastatic renal cell carcinoma (mRCC) may benefit from cytoreductive nephrectomy (CN) before targeted therapy (TT), and to assess the overall survival of patients undergoing CN and TT versus TT alone. We identified 88 patients who underwent CN at our institution prior to planned TT and 35 patients who received TT without undergoing CN. Preoperative risk factors described in the literature were assessed in our patient population (serum albumin, liver metastasis, symptomatic metastasis, clinical ≥T3 disease, retroperitoneal and supradiaphragmatic lymphadenopathy). Patients were stratified by number of pretreatment risk factors and overall survival (OS) was compared. TT patients had significantly more risk factors compared to CN patients (3.06 vs. 2.11, p<0.01). Patients who received TT alone had median OS of 5.8 months. All but one patient receiving TT alone had two or more risk factors. A comparison of the CN and TT groups was performed by constructing Kaplan-Meier curves. There was no significant difference in median OS for those patients with exactly two risk factors (447 vs. 389 days, p=0.24), and those with three or more risk factors (184 vs. 155 days, p=0.87). Using previously described pretreatment risk factors we found that patients with two or more risk factors derived no significant survival advantage from CN in the TT era. These risk factors should be incorporated in the assessment of patients for CN.


Safety of phenol vapor inhalation during performance of chemical matrixectomy to treat ingrown toenails.
Phenol is commonly used in chemical matrixectomies to treat ingrown toenails. Although many studies have documented the safety of phenol matrixectomy for the patient, few studies have investigated the procedure's safety for the physician and auxiliary health care personnel. This study was undertaken to evaluate the safety of phenol vapor inhalation for health care personnel during performance of chemical matrixectomy. The environmental concentrations of phenol in the respiration zones of the physician and an assistant were measured during an in-office surgical procedure consisting of two partial chemical matrixectomies using a 95% phenol solution. The samples were analyzed using gas chromatography with a flame ionization detector. The phenol concentration measured in the physician's respiration zone was close to Spain's established environmental limit of daily exposure, with an index of exposure of 60%. The concentration measured in the assistant's respiration zone was well below the environmental limit of daily exposure, with an index of exposure of only 10%. The performance of chemical matrixectomies is safe for health care personnel when they are exposed to the vapor of phenol solutions of up to 95% concentration for up to 21 minutes. The degree of safety would be greater with lower phenol concentrations and shorter exposure times.


Twelve-Year Follow-up After Peptide Receptor Radionuclide Therapy.
Peptide receptor radionuclide therapy (PRRT) has been used for more than 20 y as a systemic treatment approach in inoperable or metastatic somatostatin receptor-positive tumors. The purpose of this study was to analyze the long-term outcome of PRRT with regard to the most commonly used radiopharmaceuticals, 90Y-DOTATOC and 177Lu-DOTATATE. Methods: This retrospective clinical study included a total of 44 consecutive patients (27 men) with advanced tumors and enhanced somatostatin receptor expression. Mean age at initial diagnosis was 60 y (SD, 11.3 y; range, 40-84 y). Median follow-up was 80 mo. For 177Lu-PRRT, the mean number of cycles administered was 5.3 ± 2.5 and the mean activity was 27.2 ± 14.9 GBq per patient. For 90Y-PRRT, the mean number of cycles administered was 5.5 ± 2.6 and the mean activity was 14.7 ± 7.3 GBq per patient. Overall, 378 cycles were administered (mean, 8.6 ± 3.4 cycles per patient), with an overall cumulative activity of 1,514.1 GBq. Results: Median overall survival was 79 mo. Twenty-one (77.8%) of the 27 men and 9 (52.9%) of the 17 women had died 12 y after commencement of PRRT. The shortest duration of illness was 8 mo and the longest 155 mo. Severe side effects (World Health Organization grades III and IV) were seen in 9 of the 14 patients still alive. Chronic kidney disease in combination with anemia was the most common finding in the 9 patients with severe side effects. A poor prognosis was found for those patients who showed progressive disease, in comparison with patients with cumulative disease control after initial PRRT (log rank, P < 0.001), whereas women and patients with no more than 2 tumor sites seemed to especially benefit from PRRT (not reaching significance levels). Conclusion: PRRT is encouraging in terms of long-term outcome. Thirty-two percent (14/44 patients) of the patients with metastatic or inoperable disease were still alive more than 12 y after the beginning of radionuclide therapy. Possible predictors for favorable outcome are having an initial response to PRRT, having a low number of affected sites, and being female.


Sodium affects the sperm motility in the European eel.
The role of seminal plasma sodium and activation media sodium on sperm motility was examined by selectively removing the element from these two media, in European eel sperm. Sperm size (sperm head area) was also measured using an ASMA (Automated Sperm Morphometry Analyses) system, in the different conditions. Intracellular sodium [Na(+)]i was quantitatively analyzed by first time in the spermatozoa from a marine fish species. Measurement of [Na(+)]i was done before and after motility activation, by Flow Cytometry, using CoroNa Green AM as a dye. Sperm motility activation induced an increase in [Na(+)]i, from 96.72mM in quiescent stage to 152.21mM post-activation in seawater. A significant decrease in sperm head area was observed post-activation in seawater. There was a notable reduction in sperm motility when sodium was removed from the seminal plasma, but not when it was removed from the activation media. Sodium removal was also linked to a significant reduction in sperm head area in comparison to the controls. Our results indicate that the presence of the ion Na(+) in the seminal plasma (or in the extender medium) is necessary for the preservation of sperm motility in European eel, probably because it plays a role in maintaining an appropriate sperm cell volume in the quiescent stage of the spermatozoa.


Rescue of an in vitro neuron phenotype identified in Niemann-Pick disease, type C1 induced pluripotent stem cell-derived neurons by modulating the WNT pathway and calcium signaling.
Niemann-Pick disease, type C1 (NPC1) is a familial disorder that has devastating consequences on postnatal development with multisystem effects, including neurodegeneration. There is no Food and Drug Administration-approved treatment option for NPC1; however, several potentially therapeutic compounds have been identified in assays using yeast, rodent models, and NPC1 human fibroblasts. Although these discoveries were made in fibroblasts from NPC1 subjects and were in some instances validated in animal models of the disease, testing these drugs on a cell type more relevant for NPC1 neurological disease would greatly facilitate both study of the disease and identification of more relevant therapeutic compounds. Toward this goal, we have generated an induced pluripotent stem cell line from a subject homozygous for the most frequent NPC1 mutation (p.I1061T) and subsequently created a stable line of neural stem cells (NSCs). These NSCs were then used to create neurons as an appropriate disease model. NPC1 neurons display a premature cell death phenotype, and gene expression analysis of these cells suggests dysfunction of important signaling pathways, including calcium and WNT. The clear readout from these cells makes them ideal candidates for high-throughput screening and will be a valuable tool to better understand the development of NPC1 in neural cells, as well as to develop better therapeutic options for NPC1.


Cost-effectiveness of physical activity interventions in adolescents: model development and illustration using two exemplar interventions.
To develop a model to assess the long-term costs and health outcomes of physical activity interventions targeting adolescents. A Markov cohort simulation model was constructed with the intention of being capable of estimating long-term costs and health impacts of changes in activity levels during adolescence. The model parameters were informed by published literature and the analysis took a National Health Service perspective over a lifetime horizon. Univariate and probabilistic sensitivity analyses were undertaken. School and community. A hypothetical cohort of adolescents aged 16 years at baseline. Two exemplar school-based: a comparatively simple, after-school intervention and a more complex multicomponent intervention compared with usual care. Incremental cost-effectiveness ratio as measured by cost per quality-adjusted life year gained. The model gave plausible estimates of the long-term effect of changes in physical activity. The use of two exemplar interventions suggests that the model could potentially be used to evaluate a number of different physical activity interventions in adolescents. The key model driver was the degree to which intervention effects were maintained over time. The model developed here has the potential to assess long-term value for money of physical activity interventions in adolescents. The two applications of the model indicate that complex interventions may not necessarily be the ones considered the most cost-effective when longer-term costs and consequences are taken into account.


Arabidopsis transcriptome reveals control circuits regulating redox homeostasis and the role of an AP2 transcription factor.
Sensors and regulatory circuits that maintain redox homeostasis play a central role in adjusting plant metabolism and development to changing environmental conditions. We report here control networks in Arabidopsis (Arabidopsis thaliana) that respond to photosynthetic stress. We independently subjected Arabidopsis leaves to two commonly used photosystem II inhibitors: high light (HL) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Microarray analysis of expression patterns during the period of redox adjustment to these inhibitors reveals that 20% and 8% of the transcriptome are under HL and DCMU regulation, respectively. Approximately 6% comprise a subset of genes common to both perturbations, the redox responsive genes (RRGs). A redox network was generated in an attempt to identify genes whose expression is tightly coordinated during adjustment to homeostasis, using expression of these RRGs under HL conditions. Ten subnetworks were identified from the network. Hierarchal subclustering of subnetworks responding to the DCMU stress identified novel groups of genes that were tightly controlled while adjusting to homeostasis. Upstream analysis of the promoters of the genes in these clusters revealed different motifs for each subnetwork, including motifs that were previously identified with responses to other stresses, such as light, dehydration, or abscisic acid. Functional categorization of RRGs demonstrated involvement of genes in many metabolic pathways, including several families of transcription factors, especially those in the AP2 family. Using a T-DNA insertion in one AP2 transcription factor (redox-responsive transcription factor 1 [RRTF1]) from the RRGs, we showed that the genes predicted to be within the subnetwork containing RRTF1 were changed in this insertion line (Deltarrtf1). Furthermore, Deltarrtf1 showed greater sensitivity to photosynthetic stress compared to the wild type.


Prince Takamatsu memorial lecture. Rare cancers: clues to genetic mechanisms.
Virtually every kind of cancer can occur in hereditary form. Such hereditary cancers are rare, but have been informative out of proportion to their frequencies because some of them have illuminated mechanisms important in the genesis of cancer generally. The prototypes of hereditary cancer have been two uncommon tumors, retinoblastoma and Wilms' tumor. Genetic epidemiologic analysis of these tumors led to the formulation of a model for their origin following two events. According to this model the first event could already be present in the germline, thereby giving rise to the heritable forms of these diseases; the second event would occur somatically in one or more cells of the relevant organ. Both events would occur somatically in the non-hereditary form. It was proposed that the two events involved mutation or loss of both copies of a particular gene; i.e., these genes are recessive in oncogenesis. Cytogenetic analyses of rare deletion cases of retinoblastoma permitted the localization of the retinoblastoma gene to chromosomal band 13q14. Deletion was also proposed as the mechanism for the rare conjunction of Wilms' tumor and aniridia; deletions of chromosomal band 11p13 were subsequently found. Deletions were also found in tumor cells of some patients with the non-hereditary forms of these two tumors, lending support to the idea that the same loci were involved in the two forms. The application of molecular genetic techniques led to verification of the recessive hypothesis for both retinoblastoma and Wilms' tumor and to the cloning of the gene of the former. These accomplishments are leading to a new understanding of mechanisms of oncogenesis and their involvement even in the common cancers of adults.


The distribution of mRNA encoded by a new member of the neuronal nicotinic acetylcholine receptor gene family (alpha 5) in the rat central nervous system.
The cellular localization of transcripts for a new putative agonist-binding subunit of the neuronal nicotinic acetylcholine receptor (nAChR), alpha 5, was examined using in situ hybridization in the rat central nervous system (CNS), alpha 5 subunit mRNA was localized to a small number of regions when compared with two of the other known agonist-binding subunits, alpha 3 and alpha 4, alpha 5 mRNA is expressed at relatively high levels in neurons of the subiculum (pyramidal layer), presubiculum and parasubiculum (layers IV and VI), which are components of the hippocampal formation, in the substantia nigra pars compacta and ventral tegmental area, in the interpeduncular nucleus, and in the dorsal motor nucleus of the vagus nerve. Moderate hybridization signals were detected in neurons of the isocortex (layer VIb), anterior olfactory nucleus, trigeminal ganglion, superior olivary complex, nucleus of the solitary tract, and area postrema. No hybridization above background levels was seen in the amygdala, septum, thalamus, hypothalamus, or cerebellum. These results suggest that the alpha 5 subunit differs from other known agonist-binding subunits in its distribution.


Mechanistic Insights into the Nonconventional Sol-Gel Synthesis of MAX Phase M2GeC (M = V, Cr).
Despite their intriguing properties, MAX phases to date remain a class of materials overwhelmingly synthesized and studied with conventional approaches that date back to their discovery. With an ever-increasing demand for new and better materials and areas of application, developing new synthesis techniques must be at the forefront of our scientific efforts and cannot be overlooked. Sol-gel chemistry, while being a very traditional approach (especially for oxides), has so far hardly been leveraged within the MAX phase community. As a newly emerging technique to access nonoxide compounds, such as MAX phases, it offers a variety of advantages over classical solid-state chemistry, namely, milder reaction conditions and greater processibility (as previously shown for Cr2GaC). Here, the sol-gel synthesis of the two MAX phase members V2GeC and Cr2GeC, in combination with both conventional and nonconventional (microwave) heating techniques, is presented. In all instances, high yields were achieved, with only minor impurities remaining in the product. This expansion of the method to other members (apart from Cr2GaC) is a critical milestone in proving the technique's viability. Additionally, using simultaneous calorimetry and mass spectrometry, first insights into the underlying carbothermal reduction reaction are presented. Understanding the chemistry and formation mechanism will help broaden the sol-gel-based synthesis technique and increase its applicability.


Diffraction data analysis in the presence of radiation damage.
In macromolecular crystallography, the acquisition of a complete set of diffraction intensities typically involves a high cumulative dose of X-ray radiation. In the process of data acquisition, the irradiated crystal lattice undergoes a broad range of chemical and physical changes. These result in the gradual decay of diffraction intensities, accompanied by changes in the macroscopic organization of crystal lattice order and by localized changes in electron density that, owing to complex radiation chemistry, are specific for a particular macromolecule. The decay of diffraction intensities is a well defined physical process that is fully correctable during scaling and merging analysis and therefore, while limiting the amount of diffraction, it has no other impact on phasing procedures. Specific chemical changes, which are variable even between different crystal forms of the same macromolecule, are more difficult to predict, describe and correct in data. Appearing during the process of data collection, they result in gradual changes in structure factors and therefore have profound consequences in phasing procedures. Examples of various combinations of radiation-induced changes are presented and various considerations pertinent to the determination of the best strategies for handling diffraction data analysis in representative situations are discussed.


A novel alpha-thalassemia nonsense mutation in codon 23 of the alpha2-globin gene (GAG-->TAG) in a Tunisian family.
Herein we describe a novel alpha-thalassemia (thal) point mutation in the alpha2-globin gene, found in a 3-year-old Tunisian girl who had Hb Bart's (gamma4) at birth, later on presenting with moderate anemia, microcytosis and hypochromia. She had a normal Hb A2 level and no abnormal hemoglobin (Hb) fraction. After excluding most of the common Mediterranean mutations, the alpha2-globin gene was sequenced and found to have a point mutation in the heterozygous state that creates a premature stop signal for translation (GAG-->TAG or Glu-->Term) at codon 23. The same mutation was also found in the mother in the heterozygous state, while the father had a normal sequence. The presence of the mutation was also confirmed by nucleotide sequencing of the opposite strand. Since the mutation creates a restriction site for the BfaI enzyme, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based assay was established for screening purposes.


Fully Automated Analysis of Muscle Architecture from B-Mode Ultrasound Images with DL_Track_US.
B-mode ultrasound can be used to image musculoskeletal tissues, but one major bottleneck is analyses of muscle architectural parameters (i.e., muscle thickness, pennation angle and fascicle length), which are most often performed manually. In this study we trained two different neural networks (classic U-Net and U-Net with VGG16 pre-trained encoder) to detect muscle fascicles and aponeuroses using a set of labeled musculoskeletal ultrasound images. We determined the best-performing model based on intersection over union and loss metrics. We then compared neural network predictions on an unseen test set with those obtained via manual analysis and two existing semi/automated analysis approaches (simple muscle architecture analysis [SMA] and UltraTrack). DL_Track_US detects the locations of the superficial and deep aponeuroses, as well as multiple fascicle fragments per image. For single images, DL_Track_US yielded results similar to those produced by a non-trainable automated method (SMA; mean difference in fascicle length: 5.1 mm) and human manual analysis (mean difference: -2.4 mm). Between-method differences in pennation angle were within 1.5°, and mean differences in muscle thickness were less than 1 mm. Similarly, for videos, there was overlap between the results produced with UltraTrack and DL_Track_US, with intraclass correlations ranging between 0.19 and 0.88. DL_Track_US is fully automated and open source and can estimate fascicle length, pennation angle and muscle thickness from single images or videos, as well as from multiple superficial muscles. We also provide a user interface and all necessary code and training data for custom model development.


Ik6 expression provides a new strategy for the therapy of acute lymphoblastic leukemia.
Our previous study demonstrated that the dominant-negative Ikaros isoform 6 (Ik6) is overexpressed in Chinese children with newly diagnosed B-acute lymphoblastic leukemia (B-ALL) and is strongly associated with a poor outcome. The purpose of the present study was to further explore the function of Ik6 in B-ALL. The association between Ik6 expression as detected by real-time PCR and efficacy of chemotherapy was evaluated. The effect of the alteration in Ik6 on leukemic cell lines was assessed by in vitro gain-of-function and loss-of-function techniques. PCR analysis showed that Ik6 expression was decreased when patients completed induction chemotherapy and reached complete remission. Ik6 expression was significantly increased when patients suffered relapse. Stable transfection of Ik6 into the Nalm-6 cell line revealed that Ik6 enhanced proliferation of Nalm-6 cells through the promotion of G0/G1-to-S-phase transition and enhanced chemoresistance to chemotherapeutics through anti-apoptotic effects. However, Ik6 expression did not affect the invasion of Nalm-6 cells. In contrast, silencing of Ik6 in Sup-B15 cells significantly inhibited proliferation and increased chemosensitivity. The present study suggests that Ik6 may be a biological marker of chemosensitivity and relapse and Ik6 may provide a potential therapeutic strategy for ALL.


Epidemiology of hereditary neuropathy with liability to pressure palsies (HNPP) in south western Finland.
An epidemiological study of hereditary neuropathy with liability to pressure palsies (HNPP) was carried out in south western Finland, with a population of 435,000. The diagnosis was established in 69 patients from 23 unrelated families through family and medical history, clinical neurological and neurophysiological examinations and with documentation of the deletion at gene locus 17p11.2 in at least one member of each family. This gave a prevalence of at least 16/100,000, which is remarkably high. However, due to the insidious nature of HNPP, most probably it is still an underestimation. This is the first population-based prevalence figure reported for HNPP. The prevalence is somewhat lower than that obtained for CMT in the same population, which agrees with the proposal that HNPP and CMT 1A are reciprocal products of the same unequal crossing-over. The clinical pictures of our patients were, in general, similar to those previously described in HNPP.


Eimeria brunetti in epithelial cells of the bursa of Fabricius in gnotobiotic chickens.
Germ-free and conventional chickens were infected with Lactobacillus acidophilus and/or Eimeria brunetti. Germ-free chickens acquiring single infections of E. brunetti were found to have lifecycle stages of the coccidial organism in the bursa of Fabricius.


Apoptosis and P53 induction in human lung fibroblasts exposed to chromium (VI): effect of ascorbate and tocopherol.
Some forms of hexavalent chromium [Cr(VI)] are known to cause damage to respiratory tract tissue, and are thought to be human lung carcinogens. Because Cr(VI) is mutagenic and carcinogenic at doses that evoke cell toxicity, the objective of these experiments was to examine the effect of Cr(VI) on the growth, survival, and mode of cell death in normal human lung fibroblasts (HLF cells). DNA adduct formation was monitored as a marker for bioavailability of genotoxic chromium. We also examined the modulation of these endpoints by vitamins C and E. Long-term Cr(VI) exposures were employed, which decreased clonogenic cell survival by 25% to 95% in a dose-dependent manner. The predominant cellular response to Cr(VI) was growth arrest. We found that Cr(VI) caused up to 20% of HLF cells to undergo apoptosis, and documented apoptotic morphology and the phagocytosis of apoptotic bodies by neighboring cells. P53 levels increased 4- to 6-fold in chromium-treated cells. In contrast with previous studies using CHO cells, the present study using HLFs found that pretreatment with either vitamin C or E did not exhibit a significant effect on Cr-induced apoptosis or clonogenic survival. In addition, pretreatment with vitamin C did not affect the p53 induction observed after chromium treatment. Neither vitamin had any effect on Cr-DNA adduct formation. These data indicate that although pretreatment with vitamin C or E alters the spectrum of cellular and/or genetic lesions induced by chromium(VI), neither vitamin altered the initiation or progression of apoptosis in diploid human lung cells.


Categories of Intimate Partner Violence and Abuse Among Young Women and Men: Latent Class Analysis of Psychological, Physical, and Sexual Victimization and Perpetration in a UK Birth Cohort.
In the UK, around one-third of young people are exposed to Intimate Partner Violence and Abuse (IPVA) by 21 years old. However, types of IPVA victimization in this population (psychological, physical, sexual), and their relationship with impact and perpetration are poorly understood. Participants in a UK birth cohort reported IPVA victimization and perpetration by age 21. We carried out a latent class analysis, where we categorized IPVA by types/frequency of victimization, and then assigned individuals to their most probable class. Within these classes, we then estimated rates of reported: 1) types of negative impacts (sad, upset/unhappy, anxious, depressed, affected work/studies, angry/annoyed, drank/took drugs more); 2) types/frequency of perpetration. Among 2130 women and 1149 men, 32% and 24% reported IPVA victimization (of which 89% and 73% reported negative impact); 21% and 16% perpetration. Victimization responses were well represented by five classes, including three apparent in both sexes: No-low victimization (characterized by low probabilities of all types of victimization; average probabilities of women and men belonging to this class were 82% and 70%); Mainly psychological (15% and 12%); Psychological and physical victimization (4% and 7%), and two classes that were specific to women: Psychological and sexual (7%); Multi-victimization (frequent victimization for all three types; 4%). In women, all types of negative impact were most common in the Psychological and sexual and Multi-victimization classes; for men, the Psychological and physical class. In women, all types of perpetration were most common for the Mainly psychological, Psychological and physical and Multi-victimization classes; in men, the Mainly psychological and Psychological and physical classes. In this study of young people, we found categories of co-occurrence of types and frequency of IPVA victimization associated with differential rates of negative impact and perpetrating IPVA. This is consistent with emerging evidence of IPVA differentiation and its variable impact in other populations.