Proteomic investigation of metabolic shift in mammalian cell culture.
Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.


Developments in the epidemiology of stomach cancer over the past decade.
The history of stomach cancer epidemiology is reviewed. The introduction of migrant population studies in the 1960 decade that described the critical role of exposures to this disease in early life was a key event. Companion pathology studies have indicated different epidemiological patterns for 2 histological entities, intestinal and diffuse type carcinomas, and confirmed an excess of intestinal metaplasia in populations at high risk to stomach cancer. Recent results suggest that epidemiology of stomach cancer can be transformed into the epidemiology of precursor lesions, and introduction of the fiberoptic gastroscope makes technically feasible detailed studies of the relationship of precursor lesions to suspect factors, including diet, in selected geographic areas. Nitroso compounds have been identified as candidate carcinogens and the epidemiological, pathological, and chemical data display signs of internal consistency. Feeding experiments with N-methyl-N'-nitro-N-nitrosoguanidine have led to animal models that permit a coordinated epidemiological-experimental approach to stomach cancer.


Genotype-dependent differences in sleep, vigilance, and response to stimulants.
To better understand the neurobiology of sleep disorders, detailed understanding of circadian and homeostatic sleep-wake regulation in healthy volunteers is mandatory. Sleep physiology and the repercussions of experimentally-induced sleep deprivation on sleep and waking electroencephalogram (EEG), vigilance and subjective state are highly variable, even in healthy individuals. Accumulating evidence suggests that many aspects of normal sleep-wake regulation are at least in part genetically controlled. Current heritability estimates of sleep phenotypes vary between approximately 20-40 % for habitual sleep duration, to over 90 % for the spectral characteristics of the EEG in nonREM sleep. The molecular mechanisms underlying the trait-like, inter-individual variation are virtually unknown, and the human genetics of normal sleep is only at the beginning of being explored. The first studies identified distinct polymorphisms in genes contributing to the endogenous circadian clock and neurochemical systems previously implicated in sleep-wake regulation, to modulate sleep architecture and sleep EEG, vulnerability to sleep loss, and subjective and objective effects of caffeine on sleep. These insights are reviewed here. They disclose molecular mechanisms contributing to normal sleep-wake regulation in humans, and have potentially important implications for the neurobiology of sleep-wake disorders and their pharmacological treatment.


Control algorithms for artificial beta cell.
The evolution of control algorithms for closed-loop regulation of blood glucose levels is described. Because of the rapid response time of the BIOSTATOR Glucose Analyzer, a derivative algorithm can be applied to replace the previous generation of "predictor" algorithms for the calculation of dynamic insulin infusion rates.


Validation of cervical cancer screening methods in HIV positive women from Johannesburg South Africa.
HIV-infected women are at increased risk for developing cervical cancer. Women living in resource-limited countries are especially at risk due to poor access to cervical cancer screening and treatment. We evaluated three cervical cancer screening methods to detect cervical intraepithelial neoplasia grade 2 and above (CIN 2+) in HIV-infected women in South Africa; Pap smear, visual inspection with 5% acetic acid (VIA) and human papillomavirus detection (HPV). HIV-infected women aged 18-65 were recruited in Johannesburg. A cross-sectional study evaluating three screening methods for the detection of the histologically-defined gold standard CIN-2 + was performed. Women were screened for cervical abnormalities with the Digene HC2 assay (HPV), Pap smear and VIA. VIA was performed by clinic nurses, digital photographs taken and then later reviewed by specialist physicians. The sensitivity, specificity and predictive valves for CIN-2 + were calculated using maximum likelihood estimators. 1,202 HIV-infected women participated, with a median age of 38 years and CD4 counts of 394 cells/mm(3). One third of women had a high grade lesion on cytology. VIA and HPV were positive in 45% and 61% of women respectively. Estimated sensitivity/specificity for HPV, Pap smear and VIA for CIN 2+ was 92%/51.4%, 75.8%/83.4% and 65.4/68.5% (nurse reading), respectively. Sensitivities were similar, and specificities appeared significantly lower for the HPV test, cytology and VIA among women with CD4 counts ≤200 cells/mm(3) as compared to CD4 counts >350 cells/mm(3). Although HPV was the most sensitive screening method for detecting CIN 2+, it was less specific than conventional cytology and VIA with digital imaging review. Screening programs may need to be individualized in context of the resources and capacity in each area.


Correcting the F508del-CFTR variant by modulating eukaryotic translation initiation factor 3-mediated translation initiation.
Inherited and somatic rare diseases result from >200,000 genetic variants leading to loss- or gain-of-toxic function, often caused by protein misfolding. Many of these misfolded variants fail to properly interact with other proteins. Understanding the link between factors mediating the transcription, translation, and protein folding of these disease-associated variants remains a major challenge in cell biology. Herein, we utilized the cystic fibrosis transmembrane conductance regulator (CFTR) protein as a model and performed a proteomics-based high-throughput screen (HTS) to identify pathways and components affecting the folding and function of the most common cystic fibrosis-associated mutation, the F508del variant of CFTR. Using a shortest-path algorithm we developed, we mapped HTS hits to the CFTR interactome to provide functional context to the targets and identified the eukaryotic translation initiation factor 3a (eIF3a) as a central hub for the biogenesis of CFTR. Of note, siRNA-mediated silencing of eIF3a reduced the polysome-to-monosome ratio in F508del-expressing cells, which, in turn, decreased the translation of CFTR variants, leading to increased CFTR stability, trafficking, and function at the cell surface. This finding suggested that eIF3a is involved in mediating the impact of genetic variations in CFTR on the folding of this protein. We posit that the number of ribosomes on a CFTR mRNA transcript is inversely correlated with the stability of the translated polypeptide. Polysome-based translation challenges the capacity of the proteostasis environment to balance message fidelity with protein folding, leading to disease. We suggest that this deficit can be corrected through control of translation initiation.


Anthropometrical measurements and maternal visceral fat during first half of pregnancy: a cross-sectional survey.
Determining anthropometric measures that indicate different fat deposits can be useful to predict metabolic risk and set specific treatment goals, reducing negative consequences for maternal and fetal health. In cases where pre-gestational weight measure and subsequent body mass index (BMI) values cannot be determined, other anthropometric measurements may be ideal for measuring the nutritional status of pregnant women, especially in low- and middle-income countries. This study aims to identify which anthropometric measurements correlate better with the maternal fat deposits measured by ultrasound. A cross-sectional study was conducted with pregnant women from the city of Porto Alegre (city), capital of Rio Grande do Sul (state), southern Brazil, from October 2016 until January 2018. Anthropometrical variables (weight, height, mid-upper arm circumference [MUAC], circumferences of calf and neck and triceps skinfolds [TSF] and subscapular skinfolds [SBSF]), and ultrasound variables (visceral adipose tissue [VAT] and total adipose tissue [TAT]) were collected. To verify the correlation of anthropometric and ultrasound measurements, a non-adjusted and adjusted Spearman correlation was used. The study was approved by the ethics committees. The age median of the 149 pregnant women was 25 years [21-31], pre-pregnancy BMI was 26.22 kg/m² [22.16-31.21] and gestational age was 16.2 weeks [13.05-18.10]. The best measurements correlated with VAT and TAT were MUAC and SBSF, both of which showed a higher correlation than pre-pregnancy BMI. It is possible to provide a practical and reliable estimate of VAT and TAT from the anthropometric evaluation (MUAC or SBSF) that is low cost, efficient and replicable in an outpatient clinic environment, especially in low- and middle-income countries.


Symptoms of upper gastrointestinal dysfunction in Duchenne muscular dystrophy: case-control study.
A case-control study was undertaken to see if symptoms of upper gastrointestinal (UGI) (oropharyngeal, esophageal, and gastric) dysfunction occurred more frequently in males afflicted with Duchenne muscular dystrophy (DMD) than healthy controls. Subjects included 55 children with confirmed DMD and 55 age-matched controls without neuromuscular disease. All subjects and/or their parents responded to a standard set of questions concerning the frequency of symptoms of UGI dysfunction. Responses of the DMD and control groups were compared using the Wilcoxon signed rank test. A significantly higher percentage of DMD patients experienced nasal quality to the voice, dysphagia, choking while eating, the need to clear the throat during or after eating, heartburn, and vomiting during or after meals, than did controls. Only one symptom--heartburn--was found significantly more frequently in the 33 nonambulatory than the 22 ambulatory DMD subjects. These findings document that feeding difficulty and symptoms consistent with oropharyngeal, esophageal, and gastric dysfunction are more frequent in the DMD population than healthy, age-matched controls.


Tailoring of Polarizing Agents in the bTurea Series for Cross-Effect Dynamic Nuclear Polarization in Aqueous Media.
A series of 18 nitroxide biradicals derived from bTurea has been prepared, and their enhancement factors ɛ ((1)H) in cross-effect dynamic nuclear polarization (CE DNP) NMR experiments at 9.4 and 14.1 T and 100 K in a DNP-optimized glycerol/water matrix ("DNP juice") have been studied. We observe that ɛ ((1)H) is strongly correlated with the substituents on the polarizing agents, and its trend is discussed in terms of different molecular parameters: solubility, average e-e distance, relative orientation of the nitroxide moieties, and electron spin relaxation times. We show that too short an e-e distance or too long a T1e can dramatically limit ɛ ((1)H). Our study also shows that the molecular structure of AMUPol is not optimal and its ɛ ((1)H) could be further improved through stronger interaction with the glassy matrix and a better orientation of the TEMPO moieties. A new AMUPol derivative introduced here provides a better ɛ ((1)H) than AMUPol itself (by a factor of ca. 1.2).


Outcome and Safety of Insulin in the Management of Smell Loss: A Systematic Review.
Background: Olfactory dysfunction can negatively impact the quality of life and increase the risk of danger from warning odors. Various factors can cause olfactory dysfunction, including COVID-19 infection, which has increased anosmia cases. No medications are approved; however, recent studies have suggested that intranasal insulin could effectively treat olfactory dysfunction. Aim: To comprehensively evaluate the effectiveness and safety of intranasal insulin in treating olfactory dysfunction. Materials and Methods: PubMed, Google Scholar, and CENTRAL databases were searched using specific keywords, and the outputs were screened using the Rayyan AI system. Original research articles published without time frame limitations that reported the relevant outcomes were included. The reviewers performed the screening and data extraction, and any disagreements were resolved by a third senior author. Results: This systematic review identified 66 references from 3 databases, with 45 articles meeting the criteria for review after duplicates were removed. Six articles met the inclusion criteria and were selected, including 177 subjects. The selected studies consisted of various study designs, including pilot studies, placebo-controlled trials, crossover studies, and randomized clinical trials. The findings showed that intranasal insulin therapy had beneficial effects on olfactory function. Specifically, improvements were observed in olfactory detection and discrimination in patients with post-COVID-19 anosmia, olfactory threshold performance in healthy participants, and odor identification in hyposmic patients. However, there were variations in the observed effects based on different doses of insulin administered and gender differences. It also shows that using insulin provides good outcomes. Using it intranasally was safe and did not cause any complications. Conclusion: Intranasal insulin has shown promising results as a potentially safe treatment for olfactory dysfunction. Studies suggest that it can improve olfactory thresholds. Further studies are needed to investigate optimal doses and potential gender differences in response.


From Beta cell replacement to beta cell regeneration: implications for antidiabetic therapy.
Diabetes is affecting more than 25.8 million people in the United States, causing huge burden on the health care system and economy. Insulin injection, which is the predominant treatment for diabetes, is incapable of replenishing the lost insulin-producing beta cell in patients. Restoring beta cell mass through replacement therapy such as islet transplantation or beta cell regeneration through in vitro and in vivo strategies has attracted particular attentions in the field due to its potential to cure diabetes. In the aspect of islet transplantation, gene therapy, stem cell therapy, and more biocompatible immunosuppressive drugs have been tested in various preclinical animal models to improve the longevity and function of human islets against the posttransplantation challenges. In the islet regeneration aspect, insulin-producing cells have been generated through in vitro transdifferentiation of stem cells and other types of cells and demonstrated to be capable of glycemic control. Moreover, several biomarkers including cell-surface receptors, soluble factors, and transcriptional factors have been identified or rediscovered in mediating the process of beta cell proliferation in rodents. This review summarizes the current progress and hurdles in the preclinical efforts in resurrecting beta cells. It may provide some useful insights into the future drug discovery for antidiabetic purposes.


[Polycystic ovary syndrome, metabolic syndrome, cardiovascular risk and the role of insulin sensitizing agents].
The Polycystic Ovary Syndrome (PCOS) affects 6 to 10% of women of childbearing age. Insulin resistance and hyperinsulinemia are present in nearly all PCOS patients and play a central role in the development of both hyperandrogenism and metabolic syndrome (MS). MS occurs in approximately 43% of PCOS patients, raising the cardiovascular risk to up seven fold in these patients. Several serum, functional and structural markers of endothelial dysfunction and subclinical atherosclerosis were described in PCOS patients, even those young and non-obese. However, despite the fact that PCOS adversely affects the cardiovascular profile, long-term studies did not demonstrate a consistent raise in cardiovascular mortality, which seems to be more observed in the post-menopausal period. Recently, oral contraceptives are being substituted for insulin sensitizing agents (metformin and glitazones) in the PCOS treatment, due to their effects on insulin resistance and cardiovascular risk.


J.E. Purkynĕ's contributions to the physiology of the visual, the vestibular and the oculomotor systems.
J.E. Purkynĕ (1787-1869) was one of the leading physiologists in the first half of the 19th century, became the founder of the "exact subjectivism", and remained during his entire life a very careful observer in psychophysical experiments. He made moreover many important discoveries in anatomy, neuroanatomy, embryology and pharmacology. Despite his outstanding contributions to science, he remained in his general philosophy and feelings deeply attached to the Romantic spirit of the age. After Purkynĕ returned to Prague in 1849 he played an important role in his nation's search for a national identity and became a leading figure in the cultural and social life of Prague. Purkynĕ's contributions to the physiology of vision, the oculomotor and the vestibular systems are reviewed. In vision Purkynĕ discovered the Purkynĕ-tree, flicker patterns seen in diffuse light, moving light nebula in the Eigengrau, spatially patterned structures in pressure phosphenes, the antagonistic properties of Galvanic phosphenes with respect to current polarity, periodic afterimages, the Troxler effect for colour perception with the peripheral retina and the principle of the ophthalmoscope. The Purkynĕ-shift in chromatic sensitivity when the adaptation level shifts from scotopic to photopic vision was rediscovered by Purkynĕ. Purkynĕ was the first to describe the "Purkynĕ images" of the anterior and posterior surfaces of the cornea and the lens and the changes in the third Purkynĕ image when the eye accommodates. He described optokinetic nystagmus and optokinetic after-nystagmus and explained movement aftereffects as a habituation of the gaze motor system. Important contributions were made by Purkynĕ to the physiology of the vestibular system, but he did not propose the existence of peripheral receptors for this system. He believed that direct mechanical effects on certain parts of the brain (especially the cerebellum) are responsible for the mechanisms of vertigo. Purkynĕ's five categories of vertigo are still valid today (vertigo of motion, Galvanic vertigo, vertigo caused by general brain ischemia, height vertigo and vertigo caused by alcohol and other narcotics). Purkynĕ's contribution to sensory physiology can be considered as one of the essential foundations on which modern sensory physiology and psychophysics are built.


Jamaican mothers' influences of adolescent girls' sexual beliefs and behaviors.
The purpose of this study was to identify the ways in which urban Jamaican mothers influence their adolescent daughters' sexual beliefs and behaviors in order to incorporate them into the design of a family-based human immunodeficiency virus (HIV) risk reduction intervention program. Focus groups were conducted with 46 14- to 18-year-old adolescent girls and 30 mothers or female guardians of adolescent girls recruited from community-based organizations in and around Kingston and St. Andrew, Jamaica. Separate focus groups were held with mothers and daughters; each included 6 to 10 participants. Focus group sessions were scripted, led by teams that included trained Jamaican and American facilitators and note-takers, and audio-taped to ensure data accuracy. Data were analyzed using qualitative content analysis. Four major maternal influences were identified: mother-daughter relationship quality, mother-daughter sexual communication, monitoring or supervision, and maternal sexual role modeling. Mothers' and daughters' reports were consistent; both groups identified positive and negative influences within each category. Some maternal influences were positive and health promoting; others were negative and promoted unsafe sexual activity and risk for HIV and other sexually transmitted infections. These influences were incorporated into the design of a culture-specific family-based HIV risk reduction intervention tailored to the needs of urban Jamaican adolescent girls and their mothers. In order to be effective, family-based HIV risk reduction interventions should be theory based and tailored to the target audience. The four maternal influences identified in this formative study were incorporated into the subsequent intervention design.


Fetal MR lung volumetry in congenital diaphragmatic hernia (CDH): prediction of clinical outcome and the need for extracorporeal membrane oxygenation (ECMO).
Despite the ultrasound (US) based lung-to-head ratio (LHR) and first results of fetal lung volume (FLV) determination in magnetic resonance imaging (MRI), there is no reliable prenatal parameter for the clinical course and outcome of fetuses with congenital diaphragmatic hernia (CDH), in particular for the need of extracorporeal membrane oxygenation (ECMO). MR FLV measurement was evaluated in 36 fetuses with CDH using T2-weighted half-Fourier acquisition single-shot turbo spin echo (HASTE) imaging. FLV and liver herniation, respectively, were correlated with survival and the need for ECMO therapy. A total of 18 healthy fetuses served as controls. MR FLV measurement was applied to predict survival and the need for neonatal ECMO therapy and to assess liver herniation as a prognostic parameter. On MRI there was a highly significant correlation of the FLV and patients' survival (p=0.0001) and ECMO requirement, respectively (p=0.0029). Compared to normal controls mean FLV in infants who died was 10% (9.4+/-5.8 ml) and 32% in surviving infants (25+/-9.7 ml). Liver herniation significantly decreased lung volume and negatively impacted clinical outcome (p<0.0005). The MR FLV is a strong predictor of survival in CDH patients. MR FLV measurements are also valuable to identify patients who may benefit from ECMO therapy. Upward liver herniation is the most important additional prognostic parameter.


Spatial Distribution of Trehalose Dihydrate Crystallization in Tablets by X-ray Diffractometry.
Crystallization of trehalose dihydrate (C12H22O11·2H2O) was induced by storing tablets of amorphous anhydrous trehalose (C12H22O11) at 65% RH (RT). Our goal was to evaluate the advantages and limitations of two approaches of profiling spatial distribution of drug crystallization in tablets. The extent of crystallization, as a function of depth, was determined in tablets stored for different time-periods. The first approach was glancing angle X-ray diffractometry, where the penetration depth of X-rays was modulated by the incident angle. Based on the mass attenuation coefficient of the matrix, the depth of X-ray penetration was calculated as a function of incident angle, which in turn enabled us to "calculate" the extent of crystallization to different depths. In the second approach, the tablets were split into halves and the split surfaces were analyzed directly. Starting from the tablet surface and moving toward the midplane, XRD patterns were collected in 36 "regions", in increments of 0.05 mm. The results obtained by the two approaches were, in general, in good agreement. Additionally, the results obtained were validated by determining the "average" crystallization in the entire tablet by using synchrotron radiation in the transmission mode. The glancing angle method could detect crystallization up to ∼650 μm and had a "surface bias". Being a nondestructive technique, this method will permit repeated analyses of the same tablet at different time points, for example, during a stability study. However, split tablet analyses, while a "destructive" technique, provided comprehensive and unbiased depth profiling information.


Loss of CD73 shifts transforming growth factor-β1 (TGF-β1) from tumor suppressor to promoter in endometrial cancer.
In many tumors, CD73 (NT5E), a rate-limiting enzyme in adenosine biosynthesis, is upregulated by TGF-β and drives tumor progression. Conversely, CD73 is downregulated in endometrial carcinomas (EC) despite a TGF-β-rich environment. Through gene expression analyses of normal endometrium samples of the uterine cancer TCGA data set and genetic and pharmacological studies, we discovered CD73 loss shifts TGF-β1 from tumor suppressor to promoter in EC. TGF-β1 upregulated CD73 and epithelial integrity in vivo in the normal endometrium and in vitro in early stage EC cells. With loss of CD73, TGF-β1-mediated epithelial integrity was abrogated. EC cells developed TGF-β1-mediated stress fibers and macromolecule permeability, migration, and invasion increased. In human tumors, CD73 is downregulated in deeply invasive stage I EC. Consistent with shifting TGF-β1 activity, CD73 loss increased TGF-β1-mediated canonical signaling and upregulated cyclin D1 (CCND1) and downregulated p21 expression. This shift was clinically relevant, as CD73Low/CCND1High expression associated with poor tumor differentiation, increased myometrial and lymphatic/vascular space invasion, and patient death. Further loss of CD73 in CD73Low expressing advanced stage EC cells increased TGF-β-mediated stress fibers, signaling, and invasiveness, whereby adenosine A1 receptor agonist, CPA, dampened TGF-β-mediated invasion. These data identify CD73 loss as essential for shifting TGF-β activity in EC.


Asparagine-linked glycoprotein biosynthesis in rat brain: identification of glucosidase I, glucosidase II, and and endomannosidase (glucosyl mannosidase).
Previous studies from this laboratory provided evidence, largely based upon the presence of a novel alpha-D-mannosidase, suggesting that the biosynthesis of N-linked glycoproteins may be different in brain as compared to other tissues (Tulsiani, D. R. P., and Touster, O. (1985) J. Biol. Chem. 260, 13,081-13,087). In the present report we describe studies on the enzymes involved in early processing reactions. These studies indicate that the brain, like other tissues, contains glucosidases I and II. The two glucosidases were separated as distinct activities with some overlapping by chromatography on a DE-52 column. The differential inhibition studies and substrate specificity studies support our conclusion that, as in other tissues, rat brain glucosidase I cleaves alpha 1,2-linked terminal glucosyl residues, whereas glucosidase II prefers alpha 1,3-linked glucosyl residues. In addition to these two processing glucosidases, we have characterized an endo enzyme (glucosyl mannosidase) in rat brain. The endomannosidase cleaves a disaccharide (glucosyl alpha 1,3-mannose) from monoglucosylated oligosaccharides (GlcMan7-9GlcNAc). Little or no activity was observed when di- or triglucosylated oligosaccharide was used as a substrate. The pH optimum of the glucosyl mannosidase is 6.2-6.8. The enzyme appears to be an intrinsic microsomal membrane component, since washing of the microsomal membranes with salt solution did not release the enzyme in soluble form. A mixture of Triton X-100 and sodium deoxycholate is required for complete solubilization of the enzyme. The solubilized enzyme is eluted from a Bio-Gel A-1.5m column as a single peak with an apparent molecular weight of 380,000.


A prospective evaluation of C-peptide levels and colorectal adenoma incidence.
Obesity is a recognised positive risk factor for colorectal adenoma and colorectal cancer. Obesity is associated with insulin resistance and compensatory hyperinsulinaemia, and circulating insulin and C-peptide, a biomarker of insulin levels, have been positively associated with colorectal cancer risk. However, whether a similar relationship exists for colorectal adenomas, an established colorectal cancer precursor, is unclear. In a nested case-control study of 273 colorectal adenoma cases and 355 matched controls from the screening arm of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial, serum C-peptide levels were measured by a chemiluminescent immunometric assay. Multivariable unconditional logistic regression models were used to calculate odds ratios (OR) and 95% confidence intervals (CI) for colorectal adenoma within quartiles of C-peptide. Further, to explore the temporal stability of C-peptide, repeat samples from the incident adenoma cases (n=50) and controls (n=30), over a 5-year period were assayed and the intra-class correlations (ICC) estimated. In a multivariable model that included established colorectal adenoma risk factors, C-peptide levels were not associated with colorectal adenoma (Q4 vs. Q1, OR 0.83, 95% CI: 0.52-1.31; P-trend 0.32); similar null associations were observed by gender, by adenoma subsite and for advanced adenomas. Among control participants, the ICC value over a 5-year period was 0.66. Our results suggest that higher C-peptide levels were not associated with colorectal adenoma incidence in this study population. Other biological pathways associated with obesity may be more relevant to the early stages of colorectal tumorigenesis.


Long-term consequences of the absence of leptin signaling in early life.
Leptin regulates energy balance and also exhibits neurotrophic effects during critical developmental periods. However, the actual role of leptin during development is not yet fully understood. To uncover the importance of leptin in early life, the present study restored leptin signaling either at the fourth or tenth week of age in mice formerly null for the leptin receptor (LepR) gene. We found that some defects previously considered irreversible due to neonatal deficiency of leptin signaling, including the poor development of arcuate nucleus neural projections, were recovered by LepR reactivation in adulthood. However, LepR deficiency in early life led to irreversible obesity via suppression of energy expenditure. LepR reactivation in adulthood also led to persistent reduction in hypothalamic Pomc, Cartpt and Prlh mRNA expression and to defects in the reproductive system and brain growth. Our findings revealed that early defects in leptin signaling cause permanent metabolic, neuroendocrine and developmental problems.


Pollinator effectiveness is affected by intraindividual behavioral variation.
Variation in pollinator quality is fundamental to the evolution of plant-pollinator mutualisms and such variation frequently results from differences in foraging behavior. Surprisingly, despite substantial intraindividual variation in pollinator foraging behavior, the consequences for pollen removal and deposition on flowers are largely unknown. We asked how two pollen foraging behaviors of a generalist pollinator (Bombus impatiens) affect removal and deposition of heterospecific and conspecific pollen, key aspects of pollinator quality, for multiple plant species. In addition, we examined how bee body size and pollen placement among body parts shaped pollen movement. We manipulated foraging behavior types using artificial flowers, which donated pollen that captive bees then deposited on three recipient plant species. While body size primarily affected donor pollen removal, foraging behavior primarily affected donor pollen deposition. How behavior affected donor pollen deposition depended on the plant species and the quantity of donor pollen on the bee's abdomen. Plant species with smaller stigmas received significantly less pollen and fewer bees successfully transferred pollen to them. For a single plant species, heterospecific pollen interfered with conspecific pollen deposition, such that more heterospecific pollen on the bee's abdomen resulted in less conspecific pollen deposition on the flower. Thus, intraindividual variation in foraging behavior and its interaction with the amount and placement of acquired pollen and with floral morphology can affect pollinator quality and may shape plant fitness via both conspecific and heterospecific pollen transfer.


Morphological and molecular taxonomy of a new species of Fundulotrema and comments on Gyrodactylus stephanus (Monogenea: Gyrodactylidae) from Fundulus heteroclitus (Actinopterygii: Cyprinodontiformes) in Nova Scotia, Canada.
Fundulotrema porterensis n. sp. (Monogenea: Gyrodactylidae) is described from the mummichog, Fundulus heteroclitus (L.; Cyprinodontidae), inhabiting Porters Lake, Nova Scotia, Canada. The new parasite species is characterized by having a ventral bar with small anterolateral processes and linguiform membrane, differentiating it from all other known species of Fundulotrema. The morphological description of F. porterensis is supplemented with 1011 sequenced base pairs (bp) of ribosomal DNA (rDNA) spanning both internal transcribed spacers (ITS-1 and ITS-2) and 5.8S regions of the genome. A BLAST (basic local alignment search tool) search revealed that the 5.8S (157 bp) region varied by 1 bp from Gyrodactylus turnbulli Harris, 1986 and G. pictae Cable, Oosterhout, Barson and Harris, 2005, which also infect cyprinodontids. Morphometrically, F. porterensis most closely resembles Fundulotremafoxi (Rawson, 1973), but the 2 species are easily separated by length of hamuli (50.7 vs. 42.2 microm, respectively), length of anterolateral process of the ventral bar (4.9 vs. 8.9 microm), shape of marginal hooks, and shape of the ventral bar membrane. A morphological and molecular supplemental diagnosis of Gyrodactylus stephanus Mueller, 1937, from the mummichog, is also presented. This new material provides previously unrecorded information on the attributes of the ventral bar, marginal hooks, and also clarifies the structure of the male copulatory organ (MCO).


[Pituitary microadenomas - current diagnostic and treatment methods].
According to previously accepted criteria, pituitary microadenoma is characterized by a diameter of less than 10 mm. Improvement and widespread use of MRI are accompanied by increased incidence of diagnosis of these neoplasms. Pituitary microadenomas is an extremely heterogeneous group of tumors with different biological behavior, endocrine secretion and clinical symptoms despite the common MR characteristics. Treatment is mainly determined by endocrine secretion. Endocrine-active microadenoma requires medication (in case of microprolactinoma) and surgical treatment (in case of microsomatotropinoma and ACTH-releasing tumor). Follow-up is advisable for endocrine-inactive microadenoma (microincidentaloma). Modern data on the incidence, clinical and endocrine features, diagnosis and treatment of various pituitary microadenomas are discussed in the article.


Identification of functional regions in the human T-cell leukemia virus type I SU glycoprotein.
Single conservative and nonconservative amino acid substitutions were introduced into the gp45 external envelope protein (SU) of human T-cell leukemia virus type I (HTLV-I). The mutated amino acids were those identified as being conserved in HTLV-I, HTLV-II, and simian T-cell leukemia virus type I (but not in bovine leukemia virus). The mutated envelopes were tested for intracellular maturation and for function. Mutants with three major phenotypes could be defined: (i) 9 mutants with a wild-type phenotype, which included most of the conservative amino acid changes (five of seven) distributed throughout the SU protein; (ii) 8 mutants with affected intracellular maturation, 6 of which define a region in the central part of the SU protein essential for correct folding of the protein; and (iii) 13 mutants with normal intracellular maturation but impaired syncytium formation. These mutations likely affect the receptor binding step or postbinding events required for fusion. Five of these mutations are located between amino acids 75 and 101 of the SU protein, in the amino-terminal third of the molecule. The other mutations involve positions 170, 181, 195, 197, 208, 233, and 286, suggesting that two other domains, one central and one carboxy terminal, are involved in HTLV-I envelope functions.


Introduction of organised mammography screening in Tyrol: results following first year of complete rollout.
In Tyrol, Austria, the existing system of spontaneous mammography screening was switched in 2007 to an organised program by smoothly changing the established framework. This process followed most EU recommendations for organised mammography screening with the following exceptions: women aged 40-49 are part of the target population, screening is offered annually to the age group 40-59, breast ultrasound is available as an additional diagnostic tool, and double reading has not yet been implemented. After a pilot phase the program was rolled out to all of Tyrol in June 2008. The aim of this study was to analyse the performance of the organised screening system by comparing quality indices and recommended levels given in the well-established EU guidelines. Working from the results of the pilot phase, we extended the organised mammography system to all counties in Tyrol. All women living in Tyrol and covered by compulsory social insurance were invited for a mammography, in the age group 40-59 annually and in the age group 60-69 biennially. Screening mammography was offered mainly by radiologists in private practice, with further assessment performed at hospitals. Using the screening database, all well-established performance indicators were analysed and compared with accepted/desired levels as per the EU guidelines. From June 2008 to May 2009, 120,440 women were invited. Per 1000 mammograms, 14 women were recalled for further assessment, nine underwent biopsy and four cancer cases were detected. Of invasive breast cancer cases, 32.3% and 68.4% were ≤ 10 mm and ≤ 15 mm in size, respectively, and 79.2% were node-negative. The positive predictive value for further assessment and for biopsy was 25.9% and 39.9%, respectively. Estimated two-year participation rate was 57.0%. In total, 14 interval cancer cases were detected during one year of follow-up; this is 18.4% of the background incidence rate. In Tyrol, Austria, an organised mammography screening program was implemented in a smooth transition from an existing spontaneous screening system and was completely rolled out within a short time. The high level of performance already seen in the pilot phase was maintained after rollout, and improvements resulting from the pilot phase were affirmed after one year of complete rollout.


[Diagnostic value of plasma concentration of pro-brain natriuretic peptide in congestive heart failure in pediatric patients with ventricular septal defects].
The value of plasma brain natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) can reflect cardiac function and therefore can be used for diagnosing congestive heart failure (CHF) and evaluating cardiac function. There are few reports, however, on the value of BNP and NT-proBNP in pediatric cases of congenital heart defect. The aim of this study was to assess the value of plasma NT-proBNP in the diagnosis of CHF and evaluation of cardiac function in pediatric patients with ventricular septal defect (VSD). Fifty-one patients with VSD aged from 2 months to 2 years old (mean 7.9 months) were enrolled. According to the modified Ross Score, the patients were divided into three groups, no CHF group (20 patients), mild CHF group (18 patients) and moderate to severe CHF group (13 patients). Fifteen age-matched normal children were used as controls. Plasma NT-proBNP was measured using enzyme immunoassay. All patients had complete echocardiographic study, including measurement of left ventricular end diastolic volume index (LVEDVI), left ventricular end systolic wall stress (LVSEWS), heart rate corrected mean velocity of circumferential fiber shortening (mVcFc), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), and contractility index (Con). The correlation between plasma NT-proBNP level and modified Ross Score and echocardiographic cardiac functional indexes was determined. The sensitivity, specificity and ROC curve of plasma NT-proBNP for diagnosing CHF was studied. Plasma NT-proBNP was positively correlated with modified Ross Score (r = 0.75, P < 0.01). Plasma NT-proBNP concentration in moderate to severe CHF group (2061 +/- 908) fmol/ml was significantly higher than that of mild CHF group (810 +/- 335) fmol/ml, and Plasma NT-proBNP concentration in mild CHF group was higher than that in no CHF group (309 +/- 68) fmol/ml. 97.14% of normal controls and subjects in no CHF group had their plasma NT-proBNP below 400 fmol/ml. 83.3% of children in mild CHF group had their plasma NT-proBNP between (400-1400) fmol/ml while in moderate and severe CHF group 84.6% of children had their plasma NT-proBNP beyond 1400 fmol/ml. Plasma NT-proBNP was also positively correlated with LVEDVI and LVSEWS. There was no correlation among mVcFc, LVEF, LVFS, Con and plasma NT-proBNP concentration. Using plasma NT-proBNP concentration > or = 400 fmol/ml as cut-point for diagnosing CHF, the sensitivity was 89.3%, the specificity was 91.2%, and the area under the ROC curve was 0.944. Plasma NT-proBNP level could be used to assess cardiac function and diagnose CHF in pediatric patients with VSD.


[The impact of diabetes mellitus on the mortality of acute coronary syndromes].
Diabetes mellitus (DM) is a leading cause of mortality in the world, mainly on account of cardiovascular diseases. At present we know that not only DM but also other hyperglycemic states are a risk factor for coronary arterial disease. In the context of acute coronary syndromes, DM determines a worst prognosis, either in short- or long-term outcomes. Since the absolute risk of death is greater among diabetic patients when compared to non-diabetic patients, therapeutical interventions have a greater impact in terms of benefits to these patients as well. Strategies such as strict control of hyperglycemia during hospitalization, acute reperfusion management (either by thrombolysis or by percutaneous coronary intervention), use of platelet glycoprotein IIb/IIIa inhibitors and angiotensin-converting enzyme (ACE)-inhibitors have recently proven to be of greater benefit for diabetics over non-diabetic patients. Meanwhile, in spite of all proven benefits of the use of evidence-based interventions to the treatment of acute coronary syndromes on diabetic patients, there is still an under utilization of these measures. Therefore, taking into account the predictions of an increasing number of diabetics in the world for the future years, and the fact that acute coronary syndromes will be the leading cause of death among them, it becomes increasingly necessary for both cardiologists and endocrinologists to work together in order to reduce the unfavorable outcomes that are expected to arise.


Positive selection acts on regulatory genetic variants in populations of European ancestry that affect ALDH2 gene expression.
ALDH2 is a key enzyme in alcohol metabolism that protects cells from acetaldehyde toxicity. Using iHS, iSAFE and FST statistics, we identified regulatory acting variants affecting ALDH2 gene expression under positive selection in populations of European ancestry. Several SNPs (rs3184504, rs4766578, rs10774625, rs597808, rs653178, rs847892, rs2013002) that function as eQTLs for ALDH2 in various tissues showed evidence of strong positive selection. Very large pairwise FST values indicated high genetic differentiation at these loci between populations of European ancestry and populations of other global ancestries. Estimating the timing of positive selection on the beneficial alleles suggests that these variants were recently adapted approximately 3000-3700 years ago. The derived beneficial alleles are in complete linkage disequilibrium with the derived ALDH2 promoter variant rs886205, which is associated with higher transcriptional activity. The SNPs rs4766578 and rs847892 are located in binding sequences for the transcription factor HNF4A, which is an important regulatory element of ALDH2 gene expression. In contrast to the missense variant ALDH2 rs671 (ALDH2*2), which is common only in East Asian populations and is associated with greatly reduced enzyme activity and alcohol intolerance, the beneficial alleles of the regulatory variants identified in this study are associated with increased expression of ALDH2. This suggests adaptation of Europeans to higher alcohol consumption.


Topology of classical molecular optimal control landscapes for multi-target objectives.
This paper considers laser-driven optimal control of an ensemble of non-interacting molecules whose dynamics lie in classical phase space. The molecules evolve independently under control to distinct final states. We consider a control landscape defined in terms of multi-target (MT) molecular states and analyze the landscape as a functional of the control field. The topology of the MT control landscape is assessed through its gradient and Hessian with respect to the control. Under particular assumptions, the MT control landscape is found to be free of traps that could hinder reaching the objective. The Hessian associated with an optimal control field is shown to have finite rank, indicating an inherent degree of robustness to control noise. Both the absence of traps and rank of the Hessian are shown to be analogous to the situation of specifying multiple targets for an ensemble of quantum states. Numerical simulations are presented to illustrate the classical landscape principles and further characterize the system behavior as the control field is optimized.


Development and Implementation of a Corriedale Ovine Brain Atlas for Use in Atlas-Based Segmentation.
Segmentation is the process of partitioning an image into subdivisions and can be applied to medical images to isolate anatomical or pathological areas for further analysis. This process can be done manually or automated by the use of image processing computer packages. Atlas-based segmentation automates this process by the use of a pre-labelled template and a registration algorithm. We developed an ovine brain atlas that can be used as a model for neurological conditions such as Parkinson's disease and focal epilepsy. 17 female Corriedale ovine brains were imaged in-vivo in a 1.5T (low-resolution) MRI scanner. 13 of the low-resolution images were combined using a template construction algorithm to form a low-resolution template. The template was labelled to form an atlas and tested by comparing manual with atlas-based segmentations against the remaining four low-resolution images. The comparisons were in the form of similarity metrics used in previous segmentation research. Dice Similarity Coefficients were utilised to determine the degree of overlap between eight independent, manual and atlas-based segmentations, with values ranging from 0 (no overlap) to 1 (complete overlap). For 7 of these 8 segmented areas, we achieved a Dice Similarity Coefficient of 0.5-0.8. The amygdala was difficult to segment due to its variable location and similar intensity to surrounding tissues resulting in Dice Coefficients of 0.0-0.2. We developed a low resolution ovine brain atlas with eight clinically relevant areas labelled. This brain atlas performed comparably to prior human atlases described in the literature and to intra-observer error providing an atlas that can be used to guide further research using ovine brains as a model and is hosted online for public access.


New epidemiologic findings about insomnia and its treatment.
This paper examines several clinical concerns about the shorter half-life benzodiazepine hypnotics from an epidemiologic perspective. It draws on data from (1) 1979 and 1990 comprehensive probability-based U.S. national household surveys of the medical use of psychotherapeutic medications; (2) a 1990 four-city community-based volunteer call-in survey of the beneficial and adverse effects of hypnotics; and (3) an analogous random-digit dialing telephone survey in the general population. The issues addressed are abuse liability, rebound, depersonalization/derealization, paranoid feelings, accidents/injuries, and the unexamined consequences of the target illness in assessments of benefit-risk. In populations representative of everyday outpatient practice, we found that (1) the abuse liability of benzodiazepine hypnotics with shorter and longer elimination half-lives was generally low and comparable; (2) prevalence rates for rebound were low and not differential for flurazepam, temazepam, triazolam, and OTC sleeping pills; (3) reports of a single or an occasional experience involving depersonalization/derealization or paranoid feelings were fairly frequent in normals, in insomnia patients prior to treatment, and in persons with untreated insomnia; (4) treatment-emergent rates of occurrence for these same symptoms were low and not drug-specific; (5) past-year prevalence rates for serious accidents/injuries were much higher for chronic untreated insomnia than for normal controls and most groups treated with psychotherapeutic medications. A high proportion of past-year users of hypnotics were satisfied with their medication and would take it again.


Destruction of Metal-Organic Frameworks: Positive and Negative Aspects of Stability and Lability.
Metal-organic frameworks (MOFs), constructed from organic linkers and inorganic building blocks, are well-known for their high crystallinity, high surface areas, and high component tunability. The stability of MOFs is a key prerequisite for their potential practical applications in areas including storage, separation, catalysis, and biomedicine since it is essential to guarantee the framework integrity during utilization. However, MOFs are prone to destruction under external stimuli, considerably hampering their commercialization. In this Review, we provide an overview of the situations where MOFs undergo destruction due to external stimuli such as chemical, thermal, photolytic, radiolytic, electronic, and mechanical factors and offer guidelines to avoid unwanted degradation happened to the framework. Furthermore, we discuss possible destruction mechanisms and their varying derived products. In particular, we highlight cases that utilize MOF instability to fabricate varying materials including hierarchically porous MOFs, monolayer MOF nanosheets, amorphous MOF liquids and glasses, polymers, metal nanoparticles, metal carbide nanoparticles, and carbon materials. Finally, we provide a perspective on the utilization of MOF destruction to develop advanced materials with a superior hierarchy for various applications.


Ouabain induces secretion of proatrial natriuretic factor by rat atrial cardiocytes.
Neonatal rat atrial and ventricular cardiocytes in primary culture secrete proatrial natriuretic factor (proANF), the 126-amino acid precursor of atrial natriuretic factor (ANF). The cellular mechanisms of proANF secretion differ in the two cell types: atrial cardiocytes store proANF in abundant secretory granules before secretion, whereas ventricular cardiocytes secrete the precursor rapidly after synthesis. In this study, we used the Na+-K+-adenosinetriphosphatase (ATPase) inhibitor ouabain to investigate the functional significance of these differing secretory mechanisms. Ouabain increased immunoreactive ANF (iANF) secretion by atrial cardiocytes 1.5- to 2.4-fold. Metabolic labeling studies demonstrated that proANF was the form of iANF released in response to ouabain. Ventricular secretion of iANF was unaffected by the Na+-K+-ATPase inhibitor. These observations are consistent with a model in which atrial cardiocytes are able to release proANF via a regulated secretory pathway, whereas ventricular cardiocytes utilize a constitutive secretory pathway. The ability of ouabain to stimulate proANF secretion suggests that an increase in intracellular calcium concentration may promote fusion of secretory granules with atrial cell membranes thereby mediating exocytosis of stored proANF.


Adherence to breastfeeding guidelines and maternal weight 6 years after delivery.
There is a dearth of information on the long-term maternal effects of breastfeeding. The objective of this study was to examine adherence to breastfeeding recommendations of exclusive breastfeeding for ≥4 months and continuation of breastfeeding for ≥1 year and maternal weight retention 6 years after delivery. Using data from the Infant Feeding Practices Study II (IFPS II), we categorized women by the degree to which they met breastfeeding recommendations. Mothers' self-reported weight 6 years after delivery (IFPS Year 6 Follow-Up) was compared with self-reported prepregnancy weight from IFPS II. Using linear regression models, adjusting for covariates, we examined associations between breastfeeding recommendation adherence and weight retention. Of the 726 women in our study, 17.9% never breastfed. Among those who initiated breastfeeding, 29.0% breastfed exclusively for ≥4 months, and 20.3% breastfed exclusively for ≥4 months and continued breastfeeding for ≥12 months. Prepregnancy BMI modified the association between breastfeeding recommendation adherence and weight retention. Adjusting for covariates, we found no association between breastfeeding recommendations adherence and weight retention among normal and overweight mothers. Among obese mothers, there was a significant linear trend (P = .03), suggesting that those who fully adhered to breastfeeding recommendations retained less weight (-8.0 kg) than obese women who never breastfed. This study suggests that improving adherence to breastfeeding recommendations may help reduce long-term maternal weight retention among obese mothers. Larger studies, with diverse populations and similar longitudinal designs, are needed to explore this relationship.


[Clinical significance of nondestructive forms of appendicitis in children].
On the basis of an analysis of remote results of appendectomy for non-destructive forms of appendicitis in children the authors make a conclusion that appendectomy was not justified in these forms of the disease. To differentiate destructive and non-destructive forms of appendicitis the authors recommend to use electromyography and laparoscopy.


Thoracic cancer surgery during the COVID-19 pandemic: a consensus statement from the Thoracic Domain of the Asian Society for Cardiovascular and Thoracic Surgery.
Healthcare resources have been mobilized to combat the COVID-19 pandemic of 2020. The Thoracic Domain of the Asian Society for Cardiovascular and Thoracic Surgery reports a consensus statement on the provision of thoracic cancer surgery during this pandemic. A Thoracic Experts Panel was convened by the Society. A consensus on the provision, safety, and setting of thoracic cancer surgery during the pandemic was obtained through a Delphi process. Responses were received from 26 panel members (96% response rate) from 10 regions across Asia. The Society recommended that elective thoracic cancer surgery services may need to be reduced or postponed if medical resources were needed for COVID-19 patients, especially intensive care unit beds and ventilators. However, thoracic cancer surgery should proceed as normal for all solid tumors, without restrictions based on disease stage, availability of non-surgical treatment options, or patient condition (unless there is a high likelihood of postoperative intensive care unit stay). Aerosol-forming procedures should be avoided intra- and perioperatively. The surgical approach does not make a difference in terms of safety. Services for thoracic cancer patients should be offered only in hospitals that maintain isolation wards for patients with confirmed or suspected COVID-19. Services for patients with thoracic cancer should be maintained during the COVID-19 pandemic. The position of the Society is that thoracic surgeons have a responsibility to perform good surgical management of thoracic cancer during the pandemic, to advocate for patients' rights to receive it, and to safeguard patients and staff from infection.


[Children's scalds from tearing of hot water bottle].
Children scalded by the tearing of hot-water bottles presented a sudden and significa tive increase around 2000 in Chile. For this reason the Aid to Burned Children Corporation (COA-NIQUEM) publicized prevention concepts concerning the careful use of this device and raised a voice of alarm to authorities. Later, the National Normalization Institute introduced quality certification standards for hot water bottles sold in the country. Determine the impact of preventive measures and quality certification for hot water bottles in burns caused by tearing of the device on children. Review of 795 patients under 15 admitted with injuries caused by hot-water bottle rupture between 2000 and 2014 at COANIQUEM Santiago. The frecuency of burns by the ethiological agent is determined and the epidemiological profile are compared in the initial and final quinquenial period. Between 2000-2004, an increase of 272.7% was recorded and then a decrease of 81.3 % was found between 2005-2014. Demographic characteristics, month of occurrence and need of some kind of surgery were similar in both periods. The number of locations and injuries diminished, and are concentrated in one burn (77.8%). In the first period the main anatomical region affected was lower limbs and in the second period abdominal and pelvis are also frequent. An important decrease of burns by hot-water bottle rupture with significant changes in their epidemiological characteristic is verified. This coincides with preventive measures and manufacturing standards regulations and quality control.


Office hysteroscopy study in consecutive miscarriage patients.
To assess the prevalence of uterine anatomical abnormalities found by office diagnostic hysteroscopy in a population of patients experiencing more than two consecutive miscarriages and compare the prevalence of uterine abnormalities between patients with two miscarriages and those with three or more consecutive miscarriages. A cross-sectional study of 66 patients with two or more consecutive miscarriages diagnosis was conducted. Patients were divided into two groups: Group A (up to two miscarriages, 23 patients), and Group B (3 miscarriages, 43 patients). They underwent an outpatient diagnostic hysteroscopy study, with either congenital or acquired abnormalities of the uterine cavity being identified. Uterine changes were found in 22 (33.3%) patients, with 9 cases of congenital changes [arcuate uterus (4 cases), septate uterus (2 cases), and bicornuate uterus (1 case)], and 13 patients with acquired changes [intrauterine adhesions (7 cases), endometrial polyp (4 cases), and uterine leiomyoma (2 cases)]. No significant differences were found between the groups as regarding both acquired and congenital uterine changes. A positive correlation was found between anatomical changes on hysteroscopy and number of miscarriages (r = 0.31; p = 0.02). Patients with more than two miscarriages have a high prevalence of uterine cavity abnormalities diagnosed by hysteroscopy; however there are no differences in prevalence or distribution of these lesions related to the number of recurrent miscarriages.


Actual Anti-TNF Trough Levels Relate to Serum IL-10 in Drug-Responding Patients With Crohn's Disease.
Patients with Crohn's disease (CD) responding to anti-tumor necrosis factor (anti-TNF) show great variability in serum drug levels, even within the therapeutic range. We aimed at exploring the role of inflammatory, genetic, and bacterial variables in relation to anti-TNF through levels in CD patients. Consecutive CD patients receiving stable doses of infliximab or adalimumab were included. Clinical and analytical parameters were recorded. Cytokine response, bacterial DNA translocation, and several immune-related genes' genotypes were evaluated, along with serum through anti-TNF drug levels. A linear regression analysis controlled by weight and drug regimen was performed. One hundred nineteen patients were initially considered. Five patients on infliximab and 2 on adalimumab showed antidrug antibodies in serum and were excluded. One hundred twelve patients were finally included (62 on infliximab, 50 on adalimumab). Fourteen patients on infliximab and 15 on adalimumab (22.6% vs 30%, P = 0.37) were receiving an intensified drug regimen. C-reactive protein (CRP), fecal calprotectin, Crohn's Disease Activity Index, leukocyte count, and albumin levels in plasma were not significantly associated with infliximab or adalimumab levels in the multivariate analysis. Serum interleukin-10 (IL-10) levels were directly related to infliximab (Beta = 0.097, P < 0.0001) and adalimumab levels (Beta = 0.069, P = 0.0241). The best multivariate regression model explaining the variability of serum infliximab and adalimumab levels included IL-10. Predicted drug levels by this model robustly fitted with actual drug levels (R2 = 0.841 for infliximab, R2 = 0.733 for adalimumab). Serum IL-10 is significantly related to serum anti-TNF levels in CD patients, showing how the disposition of anti-TNF drugs is significantly influenced by the degree of immunological activation.


Postoperative intraspinal subdural collections after pediatric posterior fossa tumor resection: incidence, imaging, and clinical features.
Postoperative intraspinal subdural collections in children after posterior fossa tumor resection may temporarily hinder metastasis detection by MR imaging or CSF analysis, potentially impacting therapy. We investigated the incidence, imaging and clinical features, predisposing factors, and time course of these collections after posterior fossa tumor resection. Retrospective review of postoperative spine MRI in 243 children (5.5 ± 4.6 years of age) from our clinical data base postresection of posterior fossa tumors from October 1994 to August 2010 yielded 37 (6.0 ± 4.8 years of age) subjects positive for postoperative intraspinal subdural collections. Their extent and signal properties were recorded for postoperative (37/37), preoperative (15/37), and follow-up spine (35/37) MRI. Risk factors were compared with age-matched internal controls (n = 37, 5.9 ± 4.5 years of age). Associations of histology, hydrocephalus and cerebellar tonsillar herniation, and postoperative intracranial subdural collections with postoperative intraspinal subdural collections were assessed by the Fisher exact test or χ(2) test. The association between preoperative tumor volume and postoperative intraspinal subdural collections was assessed by the Wilcoxon rank sum test. The overall incidence of postoperative intraspinal subdural collections was 37/243 (15.2%), greatest ≤7 days postoperatively (36%); 97% were seen 0-41 days postoperatively (12.9 ± 11.0 days). They were T2 hyperintense and isointense to CSF on T1WI, homogeneously enhanced, and resolved on follow-up MR imaging (35/35). None were symptomatic. They were associated with intracranial subdural collections (P = .0011) and preoperative tonsillar herniation (P = .0228). Postoperative intraspinal subdural collections are infrequent and clinically silent, resolve spontaneously, and have a distinctive appearance. Preoperative tonsillar herniation appears to be a predisposing factor. In this series, repeat MR imaging by 4 weeks documented improvement or resolution of these collections in 88%.


New medical device regulations: the regulator's view.
Advances in medical device technology have been dramatic in recent years resulting in both an increased number of medical devices and an increase in the invasiveness and critical function which devices perform. Two new regulations entered into force in Europe in May 2017, the Medical Device Regulation (MDR) and the In Vitro Diagnostic Device Regulation (IVDR). These regulations will replace the current directives over the coming years. These regulations, for the first time introduce requirements relating to registries.Medical device manufacturers are required to have systematic methods for examining their devices once available on the market, by systematically gathering, recording and analysing data on safety and performance.Registries can assist public health protection in very practical ways, for example, to help urgently identify patients or devices. Registries can also be powerful tools for collecting and appraising real-world clinical evidence concerning medical devices. Clinical investigations are limited in terms of the sample size and the duration of follow-up which can reasonably be expected. Registries may also be the only available tool to examine rare adverse effects, sub-populations or for time durations which it is not possible or feasible to study in a clinical investigation. By ensuring that a core dataset is collected which can be compared to other registries or trial data, it is possible to pool data to better examine outcomes. There are a range of excellent initiatives which have aimed at ensuring the appropriate regulatory application of registry data. Cite this article: EFORT Open Rev 2019;4 DOI: 10.1302/2058-5241.4.180061.


Diabetic syndrome induced by monosodium aspartate administration in Microtus arvalis Pallas.
Administration of monosodium aspartate (MSA) to neonatal voles, Microtus arvalis Pallas induced neuronal necrosis in several brain regions including the arcuate and ventromedial hypothalamic nuclei. In the growing stage, 56.7% showed glycosuria. In the mild diabetic voles, blood glucose concentrations rose to 180 mg/dl (72 mg/dl in normal), and plasma insulin levels were also increased to 112 microU/ml (21 microU/ml in normal). In the severe diabetic voles, on the other hand, blood glucose concentrations rose over 250 mg/dl, and plasma insulin levels decreased under 10 microU/ml. Histopathological examination of the pancreatic islets revealed moderately enlarged islets, and nuclear hypertrophy and partial degranulation of B cells in the mild diabetic voles, and moderately to markedly enlarged islets, and marked degranulation, vacuolation and accumulation of glycogen granules of B cells in the severe diabetic voles. These results indicated that the diabetic syndrome in herbivorous voles was induced by administration of MSA and diabetic voles fell into marked insulin deficiency in the severe stage.


The Role of Tailored Public Health Messaging to Young Adults during COVID-19: "There's a lot of ambiguity around what it means to be safe".
The COVID-19 global incidence rate among young adults (age 19-40) drastically increased since summer 2020, and young adults were often portrayed by popular media as the "main spreader" of the pandemic. However, young adults faced unique challenges during the pandemic due to working in high-risk, low-paying essential service occupations, as well as having higher levels of financial insecurity and mental burden. This qualitative study aims to examine the attitudes and perceptions of health orders of young adults to better inform public health messaging to reach this demographic and increase compliance to public health orders. A total of 50 young adults residing in British Columbia, Canada, were recruited to participate in focus group in groups of four to six. Focus group discussions were conducted via teleconferencing. Thematic analysis revealed four major themes: 1) risks of contracting the disease, 2) the perceived impact of COVID-19, 3) responsibility of institutions, 4) and effective public health messaging. Contrary to existing literature, our findings suggest young adults feel highly responsible for protecting themselves and others. They face a higher risk of depression and anxiety compared to other age groups, especially when they take on multiple social roles such as caregivers and parents. Our findings suggest young adults face confusion due to inconsistent messaging and are not reached due to the ineffectiveness of existing strategies. We recommend using evidence-based strategies proven to promote behaviour change to address the barriers identified by young adults through tailoring public health messages, specifically by using positive messaging, messaging that considers the context of the intended audiences, and utilizing digital platforms to facilitate two-way communication.


[Distribution of butterflies (Lepidoptera: Hesperioidea and Papilionoidea) from Mexico State, Mexico].
The State of Mexico is a region with great biological diversity, owing to its geographical and ecological features. Regarding Hesperioidea and Papilionoidea, 15% of the Mexican species are recorded in the State of Mexico, 17% of which are endemic to the country. A checklist of the two superfamilies for the State of Mexico was integrated, based on published literature and databases at the Museo de Zoología of the Facultad de Ciencias, UNAM. The checklist is composed by six families, 22 subfamilies, 197 genera and 325 species (95 Hesperiidae, 19 Papilionidae, 35 Pieridae, 54 Lycaenidae, 20 Riodinidae, and 102 Nymphalidae). A list of each species is presented, including collecting localities, flight month, and whether data correspond to scientific collection records or literature.


Causes of unplanned hemodialysis initiation.
Half of patients starting chronic hemodialysis used a transient vascular catheter as a vascular access (unplanned initiation). An objective of the Quality Management Group of the Spanish Society of Nephrology is to achieve that 80% of the patients starting hemodialysis do it with an arteriovenous fistula. We want to review the causes of non-planned hemodialysis nowadays. In 2010, 43 patients had started chronic hemodialysis in the Hospital Ramón y Cajal in Madrid (Spain). Mean age was 61 years, 79% were men, the most frequent cause of chronic renal disease was the diabetes (23%) and Charlson Comorbidity Index was 6.3 ± 2.6. The unplanned hemodialysis occurred in 20 patients (47%), without any differences with the 23 patients who began planned hemodialysis, in none of the clinical or demographic parameters analyzed. The main cause of unplanned hemodialysis was the acute exacerbation of chronic kidney disease stage 3 or 4, previously stable, secondary to an unforeseeable intercurrent process (8 patients, 40% of the cases). One patient began after a non-recovery acute renal failure and in other 6 patients, the reason of unplanned hemodialysis initiation was not attributable to the operation Health System (in 3 cases unknown kidney chronic disease and in the other 3 cases it was patient´s responsibility). Only in 5 cases (25%), the cause could be corrigible. Most causes of unplanned hemodialysis does not come from the healthcare organization and therefore not easy to resolve it. Consequently, the objective of the Quality Group will be difficult to be achieved.


Posttraumatic scar revision: a review and case presentation.
Scar revision is a well-established procedure, but the achievement of satisfying long-term results may present a challenge. An appropriate initial management of wounds is of importance, since it has a role in determining the degree of revision required postoperatively. In addition to the conventional treatment and maturation of the scar tissue, a combination of procedures are now available which may alter the appearance of the final scar. Scar revision, followed by wound care that consists of silastic sheeting, steroid injection, and laser skin resurfacing with carbon dioxide laser (CO2), may be used as adjuncts to achieve camouflage of facial scars. Two case reports are presented to document the procedure, followed by treatment evaluation and protocol.


The DEBIT trial: an intervention to reduce antipsychotic polypharmacy prescribing in adult psychiatry wards - a cluster randomized controlled trial.
Clinical guidelines advise against prescribing more than one antipsychotic with limited exceptions. Despite this, surveys continue to report high antipsychotic polypharmacy rates. The aim of the study was to investigate the effectiveness of a multi-faceted intervention in reducing prescribing of antipsychotic polypharmacy on general adult psychiatry wards, compared with guidelines alone. A pragmatic cluster randomized controlled trial recruited 19 adult psychiatric units (clusters) from the South West of England. Participants were all ward doctors and nurses. The multi-faceted intervention comprised: an educational/CBT workbook; an educational visit to consultants; and a reminder system on medication charts. The odds of being prescribed antipsychotic polypharmacy in those patients prescribed antipsychotic medication was significantly lower in the intervention than control group when adjusted for confounders (OR 0.43, 95% CI 0.21-0.90, p=0.028). There was considerable between-unit variation in polypharmacy rates and in the change in rates between baseline and follow-up (5 months after baseline). The intervention reduced levels of polypharmacy prescribing compared to guidelines alone although the effect size was relatively modest. Further work is needed to elicit the factors that were active in changing prescribing behaviour.


Epidemiologic and Clinical Features of Mpox in Adults Aged >50 Years - United States, May 2022-May 2023.
During May 2022-May 2023, approximately 30,000 mpox cases were reported in the United States, predominantly among young adult men. Persons aged >50 years might experience more severe mpox disease because of a higher prevalence of comorbidities. Conversely, they could have residual protection from childhood smallpox vaccination against monkeypox virus infection and severe mpox, as has been suggested by investigation of some previous mpox outbreaks. To examine the characteristics of mpox cases among adults aged >50 years, analysts compared mpox epidemiology and clinical outcomes among all adults aged ≥18 years, by age group. Further, outcomes were compared among adults aged >50 years by JYNNEOS vaccination status. During May 10, 2022-May 17, 2023, among 29,984 adults with probable or confirmed mpox reported to CDC, 2,909 (9.7%) were aged >50 years, 96.3% of whom were cisgender men. Compared with adults aged 18-50 years, adults aged >50 years had higher prevalences of immunocompromising conditions (p<0.001) and HIV infection (p<0.001). Among adults with mpox aged >50 years, 27.6% had received JYNNEOS vaccination; this group had lower prevalences of constitutional symptoms (p<0.001), pruritus (p<0.001), and hospitalization (p = 0.002) compared with those who had not received JYNNEOS vaccine. Currently recommended JYNNEOS vaccination among all adults at risk for mpox should be encouraged, irrespective of childhood smallpox vaccination status.


Sentinel lymph node biopsy in head and neck squamous cell carcinoma.
Sentinel lymph node biopsy is a minimally invasive method to stage the regional lymphatics that has revolutionized the management of patients with intermediate-thickness cutaneous melanoma. Head and neck surgeons have been encouraged by the accuracy of sentinel lymph node biopsy in cutaneous melanoma and have applied the technique to patients with head and neck squamous cell carcinoma (HNSCC). The objectives of the study were 1) to study the feasibility and accuracy of sentinel lymph node biopsy as a method to stage the regional lymphatics in HNSCC and 2) to determine whether there are qualitative differences between the cutaneous and mucosal lymphatics that would affect the technique used in HNSCC. Two methods of investigation were employed: a prospective laboratory study using a feline model for sentinel lymph node biopsy and a retrospective review of patients who received lymphoscintigraphy before neck dissection and intraoperative identification of the sentinel lymph node. Lymphoscintigraphy and a gamma probe were used in four felines to study the kinetics of technetium-labeled sulfa colloid (Tc-SC) in the mucosal lymphatics. In the second part of the feline study, eight subjects were studied intraoperatively. Tc-SC and isosulfan blue dye were used to study the injection technique for the mucosal lymphatics and to determine the time course of the dye and Tc-SC to the sentinel lymph node. In Part II of the present study, a retrospective review of 33 patients with HNSCC was conducted. Twenty patients (stage N0) whose treatment included elective neck dissection were studied with preoperative lymphoscintigraphy and underwent intraoperative identification of the sentinel lymph node to determine the accuracy and feasibility of sentinel lymph node biopsy. Eight patients with palpable neck disease and five patients with recurrent or second primary disease whose previous treatment included neck dissection were also studied with lymphoscintigraphy before neck dissection. In the feline study, both Tc-SC and isosulfan blue dye traversed the lymphatics rapidly, appearing in the sentinel lymph node in less than 5 minutes. Modification of the injection technique used for cutaneous melanoma was required to depict the sentinel lymph node of the base of tongue. In the human study, the sentinel lymph node was accurately identified in 19 of 20 (95%) N0 patients. On average, 2.9 sentinel lymph nodes (range, 1-5) were identified in 2.2 (range, 1-4) levels of the neck. Sentinel lymph nodes were bilateral in 4 of 19 patients. When the sentinel lymph node was identified, it accurately predicted the pathological nodal status of the regional lymphatics. Three of 20 patients had cervical metastases, and the sentinel lymph node was identified in 2 of 3 patients with pathologic nodes (pN+). Focal areas of radiotracer uptake were identified in seven of eight patients with palpable disease. These areas corresponded to the level with palpable disease in four patients. The lymphatics delineated by lymphoscintigraphy in the five patients with previous neck dissection were outside the levels that had been dissected. Lymphoscintigraphy depicted collateral patterns of lymphatic drainage. Sentinel lymph node biopsy is technically feasible and is a promising, minimally invasive method for staging the regional lymphatics in patients with stage N0 HNSCC. Lymphoscintigraphy alone may determine the levels that require treatment in patients with disrupted or previously operated cervical lymphatics.


Management of acute acromioclavicular joint dislocations: current concepts.
Acromioclavicular joint (ACJ) injuries represent a common injury to the shoulder girdle. In the management algorithm of acute ACJ injuries complete radiological evaluation represents the key to a successful therapy. According to the classification of Rockwood the presence of a horizontal component in addition to vertical instability has to be detected. Using axillary functional views or Alexander views dynamic horizontal ACJ instability can be diagnosed in a simple, efficient and cost-effective manner reducing the number of mis-/underdiagnosed ACJ injuries. MRI should not be the imaging modality of first choice. The treatment of ACJ dislocations must consider two aspects. In addition to the correct type of injury therapy strategies should be adapted to patient's demands and compliance. Low grade AC injuries types I and II are treated non-operatively in terms of "skilful-neglect". High-grade injuries types IV-VI should be treated operatively within a time frame of 2-3 weeks after injury. A certain debate is still ongoing regarding type III injuries. Out from the literature, non-operative treatment of type III injuries results to provide at least equal functional outcomes as compared to surgical treatment associated with less complications and earlier return to professional and sports activities. If surgical treatment is indicated, open surgical procedures using pins, PDS-slings or hook plates are still widely used concurring with recently raising minimally invasive, arthroscopic techniques using new implants designed to remain in situ. Combined coracoclavicular and acromioclavicular repair are gaining in importance to restore horizontal as well as vertical ACJ stability.


Variations in the lipid peroxidation and antioxidant biomarkers in some tissues of anadromous cyprinid fish during migration.
Alburnus tarichi is an endemic fish species inhabiting the Lake Van basin of Turkey. This anadromous cyprinid fish migrates for spawning to the freshwater inlets pouring into Lake Van, which has highly alkaline and brackish water. During the migration, the fish moves to a new habitat from a different habitat and encounters many challenges. The objective of the present study is to investigate whether antioxidant responses occur in the tissues of A. tarichi during its anadromous migration. To this end, fish were sampled at three different points and in two different periods from the migration route, including in Lake Van at prespawning, at the entrance of a freshwater stream (Karasu) at spawning and in the upstream of the freshwater stream at spawning. Malondialdehyde (MDA) content and antioxidant defenses including activities of the enzymes superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and catalase and the level of glutathione were assayed in liver, gill, white muscle, trunk kidney and anterior intestine tissues. Our results showed increased MDA levels in liver, white muscle and anterior intestine and tissue-specific antioxidant responses in the freshwater environment. This study reports that alterations occurred in the antioxidant defense system indicators in the tissues of anadromous cyprinid fish during migration and that the antioxidant defenses might reflect an important role in spawning migration that ultimately leads to accomplishment of reproductive activity.


Long-term economic consequences for women of delayed childbearing and reduced family size.
Using data from the Panel Study of Income Dynamics, this study explored the association among delayed childbearing, completed family size and several measures of the economic well-being of women age 60 and older in 1976. By retirement age women who bore their first child at age 30 or older are significantly better off economically than either average-age childbearers or the childless. Economic well-being also appears to be related to family size among late childbearers . At retirement age the delayed childbearer with only one or two children appears better off than all other women. Thus, late childbearing and small family size appear associated with the highest standard of living for these women. This study also relates the experience of this early cohort of women to that of more recent birth cohorts.


A multiscale statistical mechanical framework integrates biophysical and genomic data to assemble cancer networks.
Functional interpretation of genomic variation is critical to understanding human disease, but it remains difficult to predict the effects of specific mutations on protein interaction networks and the phenotypes they regulate. We describe an analytical framework based on multiscale statistical mechanics that integrates genomic and biophysical data to model the human SH2-phosphoprotein network in normal and cancer cells. We apply our approach to data in The Cancer Genome Atlas (TCGA) and test model predictions experimentally. We find that mutations mapping to phosphoproteins often create new interactions but that mutations altering SH2 domains result almost exclusively in loss of interactions. Some of these mutations eliminate all interactions, but many cause more selective loss, thereby rewiring specific edges in highly connected subnetworks. Moreover, idiosyncratic mutations appear to be as functionally consequential as recurrent mutations. By synthesizing genomic, structural and biochemical data, our framework represents a new approach to the interpretation of genetic variation.


Endothelial cell proliferation associated with lesions of murine systemic candidiasis.
Neovascularization is associated with tumor growth and some inflammatory diseases but has not been reported to be induced by infectious agents. In a mouse model of systemic Candida albicans infection, extensive endothelial cell proliferation was seen in the periphery of brain abscesses and in the areas of fungal pyelonephritis in the kidney. This finding is important for an understanding of the pathogenesis of fungal infections and may contribute to an analysis of the mechanisms of angiogenesis.


[Inherited disorders of bilirubin metabolism].
Jaundice in an infant or older child may reflect accumulation of either unconjugated or conjugated bilirubin and could be related to inherited bilirubin disorders. Three grades of inherited unconjugated hyperbilirubinemia are recognised in humans. This spectrum of disorders is distinguished primarily on the basis of the plasma bilirubin level, the response to phenobarbital administration, and the presence or absence of bilirubin glucoronides in bile. The enzyme responsible for the conjugation of bilirubin is the bilirubin uridine-diphosphate-glucuronosyltransferase (UGT). Mutations in the gene encoding bilirubin-UGT (UGT1A1), lead to complete or partial inactivation of the enzyme causing the rare autosomal recessively inherited conditions, Crigler-Najjar syndrome type 1 (CN-1) and type 2 (CN-2). Gilbert syndrome (GS) is due to an insertional mutation at homozygous state of the TATAA element (seven TA repeats) of UGT1A1 producing a reduced level of expression of the gene. The association of GS with haemolytic anemias, e.g., Hereditary Spherocytosis (HS) or Congenital Dyserythropoietic Anemia type 2 (CDA 2), increase the hyperbilirubinemia level and the risk of cholelithiasis. Forms of chronic conjugated hyperbilirubinemia are Dubin-Johnson syndrome, Rotor syndrome, Alagille syndrome or arteriohepatic dysplasia, Wilson disease or hepatolenticular degeneration. Liver or liver cell transplantation is the therapy in some cases.


Development and validation of a PulseNet standardized protocol for subtyping isolates of Cronobacter species.
Cronobacter (formerly known as Enterobacter sakazakii) is a genus comprising seven species regarded as opportunistic pathogens that can be found in a wide variety of environments and foods, including powdered infant formula (PIF). Cronobacter sakazakii, the major species of this genus, has been epidemiologically linked to cases of bacteremia, meningitis in neonates, and necrotizing enterocolitis, and contaminated PIF has been identified as an important source of infection. Robust and reproducible subtyping methods are required to aid in the detection and investigation, of foodborne outbreaks. In this study, a pulsed-field gel electrophoresis (PFGE) protocol was developed and validated for subtyping Cronobacter species. It was derived from an existing modified PulseNet protocol, wherein XbaI and SpeI were the primary and secondary restriction enzymes used, generating an average of 14.7 and 20.3 bands, respectively. The PFGE method developed was both reproducible and discriminatory for subtyping Cronobacter species.


[Osteoma and polyposis: an unusual association].
A case of osteoma of the ethmoidal sinuses associated with nasal polyposis was treated by endoscopic nasal surgery. The pathophysiology of both diseases is reviewed. The therapeutic strategy is described and the bibliography is reviewed.


Identification of a promiscuous T-cell epitope in Mycobacterium tuberculosis Mce proteins.
The characterization of Mycobacterium tuberculosis antigens inducing CD4(+) T-cell responses could critically contribute to the development of subunit vaccines for M. tuberculosis. Here we performed computational analysis by using T-cell epitope prediction software (known as TEPITOPE) to predict promiscuous HLA-DR ligands in the products of the mce genes of M. tuberculosis. The analysis of the proliferative responses of CD4(+) T cells from patients with pulmonary tuberculosis to selected peptides displaying promiscuous binding to HLA-DR in vitro led us to the identification of a peptide that induced proliferation of CD4(+) cells from 50% of the tested subjects. This study demonstrates that a systematic computational approach can be used to identify T-cell epitopes in proteins expressed by an intracellular pathogen.


Mutational analysis of trans-membrane helices M3, M4, M5 and M7 of the fast-twitch Ca2+-ATPase.
Mutational analysis of trans-membrane helices M3, M4, M5 and M7 of the Ca2+-ATPase revealed a novel phenotypic variant, M4 [Y295A (the one-letter symbols are used for amino acid residues throughout)], displaying an increased affinity for Pi and decreased affinity for MgATP, while retaining the ability to translocate Ca2+ ions across the endoplasmic reticulum membrane. The properties of this mutant suggest that the E1-E2 equilibrium is shifted towards E2, and indicate a key role for this aromatic residue (Y295) at the end of trans-membrane helix M4. A mutant containing three amino acid residue substitutions at the end of the seventh trans-membrane helix, M7 (F834A, F835A, T837F), showed a complete loss of ATPase activity and a reduced ability to phosphorylate with Pi, although MgATP-initiated phosphorylation was unaffected. The observation that single mutations in this cluster of residues had no effect on Ca2+ transport suggests that correct anchoring of the helix at the lipid-water interface by these aromatic residues is important in the functioning of the ATPase. Mutation of polar residues in helix M3 did not affect inhibition of the ATPase by thapsigargin, thapsivillosin A or t-butyl hydroquinone, suggesting that hydrogen-bonding partners for the essential -OH groups on these inhibitors lie elsewhere in the ATPase.


Serum Brain-Derived Neurotrophic Factor is Related to Platelet Reactivity but not to Genetic Polymorphisms within BDNF Encoding Gene in Patients with Type 2 Diabetes.
The aim of this study was to investigate the association between serum concentrations of the brain-derived neurotrophic factor (BDNF), platelet reactivity and inflammatory markers, as well as its association with BDNF encoding gene variants in type 2 diabetic patients (T2DM) during acetylsalicylic acid (ASA) therapy. This retrospective, open-label study enrolled 91 patients. Serum BDNF, genotype variants, hematological, biochemical, and inflammatory markers were measured. Blood samples were taken in the morning 2-3 h after the last ASA dose. The BDNF genotypes for selected variants were analyzed by use of the iPLEX Sequenom assay. In multivariate linear regression analysis, CADP-CT >74 sec (p<0.001) and sP-selectin concentration (p=0.03) were predictive of high serum BDNF. In multivariate logistic regression analysis, CADP-CT >74 sec (p=0.02) and IL-6 concentration (p=0.03) were risk factors for serum BDNF above the median. Non-significant differences were observed between intronic SNP rs925946, missense SNP rs6265, and intronic SNP rs4923463 allelic groups and BDNF concentrations in the investigated cohort. Chronic inflammatory condition and enhanced immune system are associated with the production of BDNF, which may be why the serum BDNF level in T2DM patients with high platelet reactivity was higher compared to subjects with normal platelet reactivity in this study.


Stay out of trouble.
Young lesbian, gay, bisexual and trans people are significantly more at risk of alcohol and drug misuse than their heterosexual counterparts. Nurses can help young patients by being aware of the issues, asking about substance use, and supporting them to make healthy choices.


Multiple transcription initiation and alternative splicing in the 5' untranslated region of the core 2 beta1-6 N-acetylglucosaminyltransferase I gene.
The glycosyltransferase core 2 beta1,6 N-acetylglucosaminyltransferase I (C2GnT I) plays an important regulatory role in the synthesis of biologically significant oligosaccharide structures. This gene is expressed in a variety of cell types, including lymphocytes and mucin-producing cells. The expression pattern of this gene suggests a complex system of regulation. To investigate the molecular regulation of this gene and locate potential promoter elements, rapid amplification of cDNA ends (RACE) analysis was used to determine the 5' ends of the C2GnT I mRNAs from a number of tissues. These experiments identified five C2GnT I mRNAs that are different in their 5' untranslated regions. The RACE cDNAs had four different 5' terminal sequences (exons A, B, D, and E'), suggesting four transcription initiation sites. One mRNA form was the result of alternative exon (exon C) utilization. These exons are spread across 60 kb of DNA on human chromosome 9, and all splice to the exon (exon F) that contains the C2GnT I coding region. Reverse transcription polymerase chain reaction experiments using primers specific for each of the four 5' end exon sequences revealed that the 5' terminal exons are differentially expressed, suggesting tissue specificity for the different 5' untranslated regions. These findings are consistent with the presence of multiple tissue-specific promoters for the C2GnT I gene.


Binding of type 1-piliated Escherichia coli to vaginal mucus.
To better understand the interactions involved in bacterial adherence and the role of mucus in the pathogenesis of urinary tract infections, we developed a system to study the binding of a recombinant Escherichia coli strain, HB101/pWRS1-17, expressing type 1 pili, to vaginal mucus collected from 28 women. Bacteria bound to differing extents to all specimens examined, and preincubation of bacteria with mannose inhibited binding by 50 to 89%. Additionally, all mucus samples showed reactivity with anti-mannose antibody, and the levels of reactivity correlated with the levels of bacterial binding, suggesting that the mannose-terminal saccharides present on these glycoproteins are the receptors for the binding of type 1-piliated bacteria. Mucus specimens collected over periods of 5 days and 12 weeks exhibited significant variation in bacterial binding, indicating temporal differences in the ability of vaginal mucus to act as a receptor for type 1-piliated E. coli. The results show that vaginal mucus can bind bacteria and may thus influence the initial attachment and subsequent colonization of the vaginal and urinary tract epithelium by E. coli.


Study of tetrahydrobiopterin and 6-methyltetrahydropterin on alcohol dependence in mice.
The literature establishes a potential link between the function of tetrahydroisoquinolines in the biological actions of alcohol and the reduced pterin cofactor needed for synthesis of certain neurotransmitters. The effect of the administration of 6-methyltetrahydropterin and tetrahydrobiopterin upon alcohol withdrawal in mice was tested. Dependence was produced by ip administration of alcohol, supplemented by alcohol delivered by a subcutaneously implanted silastic tube. Administration of the reduced pterins at relatively high doses did not reduce the magnitude of the alcohol withdrawal syndrome.


Health care resource use by patients before and after a diagnosis of chronic fatigue syndrome (CFS/ME): a clinical practice research datalink study.
Our aim was to investigate patterns of health care resource use by patients before and after a diagnosis of CFS/ME, as recorded by Clinical Practice Research Datalink (CPRD) GP practices in the UK. We used a case-control study design in which patients who had a first recorded diagnosis of CFS/ME during the period 01/01/2001 to 31/12/2013 were matched 1:1 with controls by age, sex, and GP practice. We compared rates of GP consultations, diagnostic tests, prescriptions, referrals, and symptoms between the two groups from 15 years (in adults) or 10 years (in children) before diagnosis to 10 years after diagnosis. Data were available for 6710 adult and 916 child (age <18 years) matched case-control pairs. Rates of GP consultations, diagnostic tests, prescriptions, referrals, and symptoms spiked dramatically in the year when a CFS/ME diagnosis was recorded. GP consultation rates were 50% higher in adult cases compared to controls 11-15 years before diagnosis (rate ratio (RR) 1.49 (95% CI 1.46, 1.52)) and 56% higher 6-10 years after diagnosis (RR 1.56 (1.54, 1.57)). In children, consultation rates in cases were 45% higher 6-10 years before diagnosis (RR 1.45 (1.40, 1.51)) and 62% higher 6-10 years after diagnosis (RR 1.62 (1.54, 1.70)). For adults and children, rates of tests, prescriptions, referrals, and symptoms were higher in cases compared to controls for up to 10 years before and after diagnosis. Adults and children with CFS/ME have greater health care needs than the rest of the population for at least ten years before their diagnosis, and these higher levels of health care resource use continue for at least ten years after diagnosis.


Mnt loss triggers Myc transcription targets, proliferation, apoptosis, and transformation.
Myc oncoproteins are overexpressed in most cancers and are sufficient to accelerate cell proliferation and provoke transformation. However, in normal cells Myc also triggers apoptosis. All of the effects of Myc require its function as a transcription factor that dimerizes with Max. This complex induces genes containing CACGTG E-boxes, such as Ornithine decarboxylase (Odc), which harbors two of these elements. Here we report that in quiescent cells the Odc E-boxes are occupied by Max and Mnt, a putative Myc antagonist, and that this complex is displaced by Myc-Max complexes in proliferating cells. Knockdown of Mnt expression by stable retroviral RNA interference triggers many targets typical of the "Myc" response and provokes accelerated proliferation and apoptosis. Strikingly, these effects of Mnt knockdown are even manifest in cells lacking c-myc. Moreover, Mnt knockdown is sufficient to transform primary fibroblasts in conjunction with Ras. Therefore, Mnt behaves as a tumor suppressor. These findings support a model where Mnt represses Myc target genes and Myc functions as an oncogene by relieving Mnt-mediated repression.