The individual relationship between atrial fibrillation sources from CARTOFINDER mapping and atrial cardiomyopathy: The catch me if you can trial.
Targeting individual sources identified during atrial fibrillation (AF) has been used as an ablation strategy with varying results. Aim of this study was to evaluate the relationship between regions of interest (ROIs) from CARTOFINDER (CF) mapping and atrial cardiomyopathy from late gadolinium enhancement (LGE) cardiovascular magnetic resonance imaging (CMR). Twenty consecutive patients underwent index catheter ablation for persistent AF (PERS AF). Pre-processed LGE CMR images were merged with the results from CF mapping to visualize harboring regions for focal and rotational activities. Atrial cardiomyopathy was classified based on the four Utah stages. Procedural success was achieved in all patients (n = 20, 100%). LGE CMR revealed an intermediate amount of 21.41% ± 6.32% for LA fibrosis. ROIs were identified in all patients (mean no ROIs per patient n = 416.45 ± 204.57). A tendency towards a positive correlation between the total amount of atrial cardiomyopathy and the total number of ROIs per patient (regression coefficient, β = 10.86, p = .15) was observed. The degree of fibrosis and the presence of ROIs per segment showed no consistent spatial correlation (posterior: β = 0.36, p-value (p) = .24; anterior: β = -0.08, p = .54; lateral: β = 0.31, p = 39; septal: β = -0.12; p = .66; right PVs: β = 0.34, p = .27; left PVs: β = 0.07, p = .79; LAA: β = -0.91, p = .12). 12 months AF-free survival was 70% (n = 14) after ablation. The presence of ROIs from CF mapping was not directly associated with the extent and location of fibrosis. Further studies evaluating the relationship between focal and rotational activity and atrial cardiomyopathy are mandatory.


[Applied anatomical study on approach next to erector spinae for spinal canal decompression through intervertebral foramen].
To observe and measure the approach next to the erector spinae in the thoracic and lumbar segments of the spine and adjacent anatomical structures by the topographic method, to clarify the positioning method and safe range so as to provide the anatomical basis of the approach for spinal canal decompression. Twelve formaldehyde-treated adult cadaver specimens were selected, including 6 males and 6 females with an average age of 43 years (range, 27-52 years) and with an average height of 166 cm (range, 154-177 cm). The related data of the approach at T1-S1 levels were respectively measured: the distance between the lateral edge of the erector spinae and the spinous process, the length of the approach, the angle between the approach and the horizontal plane, the size of intervertebral foramen, and the vertical distance between the segmental artery and the upper edge of the vertebrae. The distance between the lateral edge of the erector spinae and the spinous process ranged from (41.75 +/- 3.29) mm to (74.54 +/- 7.08) mm. The length of the approach ranged from (66.75 +/- 10.81) mm to (97.13 +/- 13.35) mm. The angle between the approach and the horizontal plane ranged from (38.38 +/- 6.16) degrees to (53.67 +/- 4.40) degrees. The vertical distance between the segmental artery and the upper edge of the vertebrae ranged from (9.50 +/-0.60) mm to (18.30 +/- 1.56) mm. The size of foraminal was also measured. The spinal canal could reach when iliocostalis lateral edge was used as the starting point in the lumbar segments, and longissimus lateral edge as the starting point in the thoracic segments. It was confirmed that there was enough safe space for the spinal decompression without the resection of the articular process. The approach next to the erector spinae can reach spinal canal to achieve the purpose of decompression through the intervertebral foramen. The minimally invasive approach is feasible and safe. It has the value of the operative application.


Influence of nanoparticles on the haemostatic balance: between thrombosis and haemorrhage.
Maintenance of a delicate haemostatic balance or a balance between clotting and bleeding is critical to human health. Irrespective of administration route, nanoparticles can reach the bloodstream and might interrupt the haemostatic balance by interfering with one or more components of the coagulation, anticoagulation, and fibrinolytic systems, which potentially lead to thrombosis or haemorrhage. However, inadequate understanding of their effects on the haemostatic balance, along with the fact that most studies mainly focus on the functionality of nanoparticles while forgetting or leaving behind their risk to the body's haemostatic balance, is a major concern. Hence, our review aims to provide a comprehensive depiction of nanoparticle-haemostatic balance interactions, which has not yet been covered. The synergistic roles of cells and plasma factors participating in haemostatic balance are presented. Possible interactions and interference of each type of nanoparticle with the haemostatic balance are comprehensively discussed, particularly focusing on the underlying mechanisms. Interactions of nanoparticles with innate immunity potentially linked to haemostasis are mentioned. Various physicochemical characteristics that influence the nanoparticle-haemostatic balance are detailed. Challenges and future directions are also proposed. This insight would be valuable for the establishment of nanoparticles that can either avoid unintended interference with the haemostatic balance or purposely downregulate/upregulate its key components in a controlled manner.


Mitigating Bias in Generalized Linear Mixed Models: The Case for Bayesian Nonparametrics.
Generalized linear mixed models are a common statistical tool for the analysis of clustered or longitudinal data where correlation is accounted for through cluster-specific random effects. In practice, the distribution of the random effects is typically taken to be a Normal distribution, although if this does not hold then the model is misspecified and standard estimation/inference may be invalid. An alternative is to perform a so-called nonparametric Bayesian analyses in which one assigns a Dirichlet process (DP) prior to the unknown distribution of the random effects. In this paper we examine operating characteristics for estimation of fixed effects and random effects based on such an analysis under a range of "true" random effects distributions. As part of this we investigate various approaches for selection of the precision parameter of the DP prior. In addition, we illustrate the use of the methods with an analysis of post-operative complications among n = 18, 643 female Medicare beneficiaries who underwent a hysterectomy procedure at N = 503 hospitals in the US. Overall, we conclude that using the DP priori n modeling the random effect distribution results in large reductions of bias with little loss of efficiency. While no single choice for the precision parameter will be optimal in all settings, certain strategies such as importance sampling or empirical Bayes can be used to obtain reasonable results in a broad range of data scenarios.


Impact of sample preparation on mineralogical analysis of zero-valent iron reactive barrier materials.
Permeable reactive barriers (PRBs) of zero-valent iron (Fe(0)) are increasingly being used to remediate contaminated ground water. Corrosion of Fe(0) filings and the formation of precipitates can occur when the PRB material comes in contact with ground water and may reduce the lifespan and effectiveness of the barrier. At present, there are no routine procedures for preparing and analyzing the mineral precipitates from Fe(0) PRB material. These procedures are needed because mineralogical composition of corrosion products used to interpret the barrier processes can change with iron oxidation and sample preparation. The objectives of this study were (i) to investigate a method of preparing Fe(0) reactive barrier material for mineralogical analysis by X-ray diffraction (XRD), and (ii) to identify Fe mineral phases and rates of transformations induced by different mineralogical preparation techniques. Materials from an in situ Fe(0) PRB were collected by undisturbed coring and processed for XRD analysis after different times since sampling for three size fractions and by various drying treatments. We found that whole-sample preparation for analysis was necessary because mineral precipitates occurred within the PRB material in different size fractions of the samples. Green rusts quickly disappeared from acetone-dried samples and were not present in air-dried and oven-dried samples. Maghemite/magnetite content increased over time and in oven-dried samples, especially after heating to 105 degrees C. We conclude that care must be taken during sample preparation of Fe(0) PRB material, especially for detection of green rusts, to ensure accurate identification of minerals present within the barrier system.


Do youth sports prevent pediatric obesity? A systematic review and commentary.
Sport is a promising setting for obesity prevention among youth, but little is known about whether it prevents obesity. We reviewed research comparing sport participants with nonparticipants on weight status, physical activity, and diet. Among 19 studies, we found no clear pattern of association between body weight and sport participation. Among 17 studies, we found that sport participants are more physically active than those who do not participate. We found seven studies that compared the diet of sport participants with non-participants. These studies reported that youth involved in sport were more likely to consume fruits, vegetables, and milk, and also more likely to eat fast food and drink sugar-sweetened beverages and consume more calories overall. It is unclear from these results whether sports programs, as currently offered, protect youth from becoming overweight or obese. Additional research may foster understanding about how sport, and youth sport settings, can help promote energy balance and healthy body weight.


Suppression of MyD88- and TRIF-dependent signaling pathways of Toll-like receptor by (-)-epigallocatechin-3-gallate, a polyphenol component of green tea.
Toll-like receptors (TLRs) play an important role in recognition of microbial components and induction of innate immunity. The microbial components trigger the activation of two downstream signaling pathways of TLRs; MyD88- and/or TRIF-dependent pathways leading to activation of NF-kappaB. (-)-Epigallocatechin-3-gallate (EGCG), a flavonoid found in green tea, is known to inhibit NF-kappaB activation induced by many pro-inflammatory stimuli. EGCG was shown to inhibit the activity of IKKbeta which is the key kinase in the canonical pathway for NF-kappaB activation in MyD88-dependent pathway of TLRs. However, it is not known whether EGCG inhibits TRIF-dependent pathway through which more than 70% of lipopolysaccharide (LPS)-induced genes are regulated. Therefore, we attempted to identify the molecular target of EGCG in TRIF-dependent pathways of TLR3 and TLR4. EGCG inhibited the activation of IFN regulatory factor 3 (IRF3) induced by LPS, poly[I:C], or the overexpression of TRIF. The inhibition of IRF3 activation by EGCG was mediated through the suppression of the kinase activity of TBK1. However, EGCG did not inhibit activation of IRF3 induced by overexpression of constitutively active IRF3. These results suggest that the molecular target of EGCG is TBK1 in TRIF-dependent signaling pathways of TLR3 and TLR4. Therefore, our results suggest that green tea flavonoids can modulate both MyD88- and TRIF-dependent signaling pathways of TLRs and subsequent inflammatory target gene expression.


Information Fusion of Passive Sensors for Detection of Moving Targets in Dynamic Environments.
This paper addresses the problem of target detection in dynamic environments in a semi-supervised data-driven setting with low-cost passive sensors. A key challenge here is to simultaneously achieve high probabilities of correct detection with low probabilities of false alarm under the constraints of limited computation and communication resources. In general, the changes in a dynamic environment may significantly affect the performance of target detection due to limited training scenarios and the assumptions made on signal behavior under a static environment. To this end, an algorithm of binary hypothesis testing is proposed based on clustering of features extracted from multiple sensors that may observe the target. First, the features are extracted individually from time-series signals of different sensors by using a recently reported feature extraction tool, called symbolic dynamic filtering. Then, these features are grouped as clusters in the feature space to evaluate homogeneity of the sensor responses. Finally, a decision for target detection is made based on the distance measurements between pairs of sensor clusters. The proposed procedure has been experimentally validated in a laboratory setting for mobile target detection. In the experiments, multiple homogeneous infrared sensors have been used with different orientations in the presence of changing ambient illumination intensities. The experimental results show that the proposed target detection procedure with feature-level sensor fusion is robust and that it outperforms those with decision-level and data-level sensor fusion.


Identification of mimotopes of Mycobacterium leprae as potential diagnostic reagents.
An early diagnostic test for detecting infection in leprosy is fundamental for reducing patients' sequelae. The currently used lepromin is not adequate for disease diagnosis and, so far, no antigen to be used in intradermoreaction has proved to be sensitive and specific for that purpose. Aiming at identifying new reagents to be used in skin tests, candidate antigens were investigated. Random peptide phage display libraries were screened by using antibodies from leprosy patients in order to identify peptides as diagnostic reagents. Seven different phage clones were identified using purified antibodies pooled from sera of leprosy patients. When the clones were tested with serum samples by ELISA, three of them, 5A, 6A and 1B, allowed detecting a larger number of leprosy patients when compared to controls. The corresponding peptides expressed by selected phage clones were chemically synthesized. A pilot study was undertaken to assess the use of peptides in skin tests. The intradermal challenge with peptides in animals previously sensitized with Mycobacterium leprae induced a delayed-type hypersensitivity with peptide 5A (2/5) and peptide 1B (1/5). In positive controls, there was a 3/5 reactivity for lepromin and a 4/5 reactivity of the sensitized animals with soluble extract of M. leprae. The preliminary data suggest that may be possible to develop reagents with diagnostic potential based on peptide mimotopes selected by phage display using polyclonal human antibodies.


Systemic and ocular determinants of mean ocular perfusion pressure in a population-based sample.
To investigate the associations between mean ocular perfusion pressure (MOPP) and several variables including body mass index (BMI), comorbid medical conditions and various ocular parameters in a population-based sample. Cross-sectional. Data of 2091 healthy participants from a previous population based cross-sectional study were reviewed. Inclusion criteria were adults ≥40 years of age who were screened on-site for glaucoma. Data on medical history, height, weight, systolic and diastolic blood pressures (SBP and DBP, respectively) were obtained. A basic ocular examination was performed which included intraocular pressure (Tono-Pen XL, Reichert Technologies) and central corneal thickness (Pacline pachymetry;) measurement, slit-lamp examination and non-mydriatic optic disc photography (nonmydα fundus camera, Kowa). MOPP was calculated using the formula [2/3 x (DBP + 1/3(SBP-DBP)]-IOP and low MOPP was defined as MOPP ≤45 mmHg. Mean age of the subjects was 63.04 ± 9.7 years (range: 44 and 99 years) and the majority were women (74.1%, n = 1549). Mean MOPP values in normal weight (BMI < 25), overweight (BMI = 25-29.9) and in obese individuals were 46.9 ± 9.0 mmHg, 48.6 ± 9.2 mmHg and 50.7 ± 10.0 mmHg, respectively (p < 0.001, in all pairwise comparisons). In the multivariable logistic regression analysis, migraine and IOP elevation (per 1 mmHg increment) were significantly associated with a low MOPP (OR: 2.10 and 1.22, p = 0.008 and < 0.001, respectively). On the other hand, risk of low MOPP was reduced in subjects with hypertension, and with increasing age (per 1-year increment) and BMI (per 1-unit increment) (OR: 0.15, 0.97, and 0.95, respectively, and p < 0.001 for all). Migraine and elevated IOP increase the risk of low MOPP and this may have a causal relationship with impaired optic nerve head blood flow.


Identification of a novel HLA-Cw*05 allele, Cw*0503.
HLA-Cw*05 is one of the least polymorphic subgroups of HLA-C; so far only two alleles, namely Cw*0501 and Cw*0502, have been reported. We report here the identification of a third allele, Cw*0503, in a Caucasian individual. Cw*0503 is closely related to Cw*0501 with only six nucleotide substitutions clustering over a fragment of 48 nucleotides at the beginning of exon 4. All these six substitutions at the same positions have been found only in HLA-B*44 alleles, suggesting that Cw*0503 is a result of recombination between Cw*0501 and one of B*44 alleles.


Blood oxygen depletion during rest-associated apneas of northern elephant seals (Mirounga angustirostris).
Blood gases (P(O)2, P(CO)2, pH), oxygen content, hematocrit and hemoglobin concentration were measured during rest-associated apneas of nine juvenile northern elephant seals. In conjunction with blood volume determinations, these data were used to determine total blood oxygen stores, the rate and magnitude of blood O(2) depletion, the contribution of the blood O(2) store to apneic metabolic rate, and the degree of hypoxemia that occurs during these breath-holds. Mean body mass was 66+/-9.7 kg (+/- s.d.); blood volume was 196+/-20 ml kg(-1); and hemoglobin concentration was 23.5+/-1.5 g dl(-1). Rest apneas ranged in duration from 3.1 to 10.9 min. Arterial P(O)2 declined exponentially during apnea, ranging between a maximum of 108 mmHg and a minimum of 18 mmHg after a 9.1 min breath-hold. Venous P(O)2 values were indistinguishable from arterial values after the first minute of apnea; the lowest venous P(O)2 recorded was 15 mmHg after a 7.8 min apnea. O(2) contents were also similar between the arterial and venous systems, declining linearly at rates of 2.3 and 2.0 ml O(2) dl(-1) min(-1), respectively, from mean initial values of 27.2 and 26.0 ml O(2) dl(-1). These blood O(2) depletion rates are approximately twice the reported values during forced submersion and are consistent with maintenance of previously measured high cardiac outputs during rest-associated breath-holds. During a typical 7-min apnea, seals consumed, on average, 56% of the initial blood O(2) store of 52 ml O(2) kg(-1); this contributed 4.2 ml O(2) kg(-1) min(-1) to total body metabolic rate during the breath-hold. Extreme hypoxemic tolerance in these seals was demonstrated by arterial P(O)2 values during late apnea that were less than human thresholds for shallow-water blackout. Despite such low P(O)2s, there was no evidence of significant anaerobic metabolism, as changes in blood pH were minimal and attributable to increased P(CO)2. These findings and the previously reported lack of lactate accumulation during these breath-holds are consistent with the maintenance of aerobic metabolism even at low oxygen tensions during rest-associated apneas. Such hypoxemic tolerance is necessary in order to allow dissociation of O(2) from hemoglobin and provide effective utilization of the blood O(2) store.


R-factor mediated beta-lactamase production by Haemophilus influenzae.
Production of beta-lactamase by 15 strains of Haemophilus influenzae has been investigated. All the strains produce a constitutive beta-lactamase, which readily hydrolyses penicillin G, ampicillin, and cephaloridine. The beta-lactamase produced by these strains is indistinguishable from the type-IIIa enzyme commonly found in strains of Escherichia coli. The beta-lactamase gene has been transferred from the enzyme-producing strains of Haemophilus to strains of H. parainfluenzae and a strain of E. coli.


Structural correlates of function in the "opercularis" muscle of amphibians.
This study characterizes the fine structure of the "opercularis" muscles of selected frogs and salamanders (Genera: Hyla; Desmognathus; Ambystoma). The "opercularis" muscle originates on the shoulder girdle and inserts on the opercular plate in the fenestra ovalis of the otic capsule. Each of the three genera used exhibits one of the major gross dispositions of this muscle found in amphibians. In each case the "opercularis" muscle contains large numbers of tonic fibers: 80% in Hyla; 90% in Desmognathus; 45% in Ambystoma. These fibers correspond to the class-5 tonic fibers of Smith and Ovalle (1973). The remained of the fibers in the "opercularis" correspond to those in the class-3 "phasic" of Smith and Ovalle. The muscle from which the "opercularis" is derived (levator scapulae in Hyla, cucullaris in Desmognathus) is comprised of fibers which correspond to the class-2 phasic fibers of Smith and Ovalle. The fiber composition of the "opercularis" indicates that it is constructed to sustain contraction over long periods of time. This composition is supportive of the functional role in audition proposed for the muscle by Lombard and Straughan (1974). Evidence is presented that indicates that fiber size may be body size dependent and thus is an inappropriate criterion of fiber type identification.


Metabolism of [14C]glucose by postimplantation mouse embryos in vitro.
Carbon dioxide and lactate production from [14C]glucose were measured for post-implantation mouse embryos aged 6 to 9 1/2 days post coitum in static cultures with a defined medium. The rate of metabolism increased rapidly and paralleled the increase in protein content indicating a fairly uniform rate of metabolism throughout the period. At all stages studied more than 90% of the glucose utilized was converted to lactate. Over a quarter of carbon dioxide produced was derived from the C-1 position resulting in high C-1:C-6 ratios, indicating that the Pentose Phosphate Shunt is a major oxidative pathway. The influence of various culture condition on CO2 production showed that high concentrations of glucose did not affect glucose utilization whilst high lactate concentrations had a significant inhibitory effect. Pyruvate had no discernible effect.


Polysaccharide immunomodulators as therapeutic agents: structural aspects and biologic function.
Polysaccharide immunomodulators were first discovered over 40 years ago. Although very few have been rigorously studied, recent reports have revealed the mechanism of action and structure-function attributes of some of these molecules. Certain polysaccharide immunomodulators have been identified that have profound effects in the regulation of immune responses during the progression of infectious diseases, and studies have begun to define structural aspects of these molecules that govern their function and interaction with cells of the host immune system. These polymers can influence innate and cell-mediated immunity through interactions with T cells, monocytes, macrophages, and polymorphonuclear lymphocytes. The ability to modulate the immune response in an appropriate way can enhance the host's immune response to certain infections. In addition, this strategy can be utilized to augment current treatment regimens such as antimicrobial therapy that are becoming less efficacious with the advent of antibiotic resistance. This review focuses on recent studies that illustrate the structural and biologic activities of specific polysaccharide immunomodulators and outlines their potential for clinical use.


COX-2 overexpression as a biomarker of early cervical carcinogenesis: a pilot study.
Recent studies have demonstrated that cyclooxygenase-2 (COX-2) expression is up-regulated in a number of cancers. Selective inhibition of COX-2 offers a potential pharmacological strategy for cancer prevention. The COX-2 isoform is induced in response to inflammatory factors and is expressed in premalignant lesions, including cervical tissues. Few studies have investigated COX-2 expression as a biomarker for early cervical carcinogenesis. In this preliminary study, we assessed the variability of COX-2 overexpression in cervical premalignant lesions. Fifty-two patients were recruited and consented. Paired abnormal and control (normal) cervical biopsies were obtained and evaluated for high-risk human papillomavirus (HPV), inflammation, histopathological diagnosis, and COX-2 protein concentration by ELISA. Paired Student's t-test and general linear regression models were used to compare mean COX-2 protein concentrations among biopsy samples and selected risk variables. Forty-seven of fifty-two paired biopsies were evaluated. COX-2 protein concentrations were 4.9-fold greater in abnormal biopsies (CIN 1 and CIN 2) than normal biopsies. COX-2 was also significantly increased in inflammation-positive biopsies. No significant association was found between COX-2 levels and HPV high-risk positivity, age, parity, STI history, or hormonal contraceptive use, but the sample size was small. These results suggest that COX-2 induction begins in the premalignant phase of cervical carcinogenesis and is correlated with inflammation. A trial using a much larger number of specimens will allow further development of our understanding of COX-2 as a biomarker for use in chemoprevention trials.


Individual-based multiple-unit dissimilarity: novel indices and null model for assessing temporal variability in community composition.
Beta-diversity was originally defined spatially, i.e., as variation in community composition among sites in a region. However, the concept of beta-diversity has since been expanded to temporal contexts. This is referred to as "temporal beta-diversity", and most approaches are simply an extension of spatial beta-diversity. The persistence and turnover of individuals over time is a unique feature of temporal beta-diversity. Nakadai (2020) introduced the "individual-based beta-diversity" concept, and provided novel indices to evaluate individual turnover and compositional shift by comparing individual turnover between two periods at a given site. However, the proposed individual-based indices are applicable only to pairwise dissimilarity, not to multiple-temporal (or more generally, multiple-unit) dissimilarity. Here, individual-based beta-diversity indices are extended to multiple-unit cases. In addition, a novel type of random permutation criterion related to these multiple-unit indices for detecting patterns of individual persistence is introduced in the present study. To demonstrate the usage the properties of these indices compared to average pairwise measures, I applied them to a dataset for a permanent 50-ha forest dynamics plot on Barro Colorado Island in Panama. Information regarding "individuals" is generally missing from community ecology and biodiversity studies of temporal dynamics. In this context, the methods proposed here are expected to be useful for addressing a wide range of research questions regarding temporal changes in biodiversity, especially studies using traditional individual-tracked forest monitoring data.


Cerebral Microbleeds and White Matter Hyperintensities are Associated with Cognitive Decline in an Asian Memory Clinic Study.
Cerebral Small Vessel Disease (SVD); lacunes, Cerebral Microbleeds (CMBs), and White Matter Hyperintensities (WMH) have a vital role in cognitive impairment and dementia. SVD in lobar location is related to cerebral amyloid angiopathy, whereas SVD in a deep location with hypertensive arteriopathy. It remains unclear how different locations of SVD affect long-term cognitive decline. The present study aimed to analyse the association between different locations and severity of SVD with global and domain-specific cognitive decline over the follow-up interval of 3 years. We studied 428 participants who had performed MRI scans at baseline and at least 3 neuropsychological assessments. Locations of lacunes and CMBs were categorized into strictly lobar, strictly deep and mixed-location, WMH volume into anterior and posterior. The National Institute of Neurological Disorders and Stroke-Canadian Stroke Network Harmonization Neuropsychological Battery was used to assess cognitive function. To analyse the association between baseline location and severity of SVD with cognitive decline, linear regression models with generalized estimated equations were constructed to calculate the mean difference, 95% confidence interval and two-way interaction factor between time and SVD. Increased numbers of baseline CMBs were associated with a decline in global cognition as well as a decline in executive function and memory domains. Location-specific analysis showed similar results with strictly lobar CMBs. There was no association with strictly deep and mixed-location CMBs with cognitive decline. Baseline WMH volume was associated with a decline in global cognition, executive function and memory. Similar results were obtained with anterior and posterior WMH volumes. Lacunes and their locations were not associated with cognitive decline. Strictly lobar CMBs, as well as WMH volume in anterior and posterior regions, were associated with cognitive decline. Future research focuses are warranted to evaluate interventions that may prevent cognitive decline related to SVD.


Purified bacteriophage lambda O protein binds to four repeating sequences at the lambda replication origin.
The bacteriophage lambda O protein is needed for initiation of lambda DNA replication. Several lines of evidence suggest that initiation requires that this protein interacts with a specific sequence called ori (for origin) in lambda DNA. We have purified this protein to near homogeneity and studied the protection against nuclease cleavage of the origin DNA sequences. Our data demonstrate that the O protein binds within an interval of about 95 base pairs (bp), which contains four tandemly arranged 19bp repeating sequences, ATCCCTCAAAACGA (G)GG GAT(A). At a low concentration of O protein, the inner two repeats are primarily covered, while binding to the outer two repeats requires a high concentration of O protein. From the molecular size of O protein (32,000 daltons), and the internal symmetry in each 19bp repeat, we inferred that the O protein may bind in dimeric form, and that the 95bp region may be filled only when four such dimers have bound. This interaction is discussed in connection with the "activation" of the ori by O protein leading to initiation of DNA synthesis.


[Dependence of acoustic-motor reaction of healthy individuals from geomagnetic activity].
During February-April, 2008 using special computer test, a daily monitoring of simple acoustic-motor reaction was carried out in 18 healthy tested individuals. We found a significant decrease in the speed of acoustic-motor reaction the day before and the same day geomagnetic disturbance occurred, as well as the same and 2-3 days after a geomagnetic calm occurred. Presumably, either an essential increase or a decreases of geomagnetic activity are adverse factors for the functional state of a central nervous system.


H2 production by Selenomonas ruminantium in the absence and presence of methanogenic bacteria.
Selenomonas ruminantium is a nonsporeforming anaerobe that ferments carbohydrates primarily to lactate, propionate, acetate and CO2. H2 production by this species has not been previously reported. We found, however, that some strains produce trace amounts of H2 which can be detected by sensitive gas chromatographic procedures. H2 production is increased markedly, in some cases almost 100-fold, when the selenomonads are co-cultured with methane-producing bacteria. Growth of the methane-producing bacteria depends on H2 production by the selenomonads and the subsequent use of H2 for the reduction of CO2 to CH4. Although no free H2 accumulates in the mixed cultures, the amount of H2 formed by the selenomonads can be calculated from the amount of methane produced. These studies indicate that the conventional methods for measuring H2 production by pure cultures do not provide an adequate estimate of an organism's potential for forming H2 in an anaerobic ecosystem when H2 is rapidly used, e.g., for formation of CH4.


RACK1 binds to Smad3 to modulate transforming growth factor-beta1-stimulated alpha2(I) collagen transcription in renal tubular epithelial cells.
Although it is clear that transforming growth factor-beta1 (TGF-beta1) is critical for renal fibrogenesis, the complexity of the involved mechanisms is increasingly apparent. TGF-beta1 stimulates phosphorylation of Smad2/3 and activates other signaling molecules as well. The molecular link between these other kinases and Smads is not known. We sought new binding partners for Smad3 in renal cells and identified receptor for activated protein kinase C 1 (RACK1) as a novel binding partner of Smad3. The linker region of Smad3 and the tryptophan-aspartic acid repeat 6 and 7 of RACK1 are sufficient for the association. RACK1 also interacts with Smad3 in the human kidney epithelial cell line, HKC. Silencing RACK1 increases transcriptional activity of TGF-beta1-responsive promoter sequences of the Smad binding element (SBE), p3TP-Lux, and alpha2(I) collagen. Conversely, overexpressed RACK1 negatively modulates alpha2(I) collagen transcriptional activity in TGF-beta1-stimulated cells. RACK1 did not affect phosphorylation of Smad3 at the C terminus or in the linker region. However, RACK1 reduced direct binding of Smad3 to the SBE motif. Mutating a RACK1 tyrosine at residue 246, but not at 228, decreased the inhibitory effect of RACK1 on both alpha2(I) collagen promoter activity and Smad binding to SBE induced by TGF-beta1. These results suggest that RACK1 modulates transcription of alpha2(I) collagen by TGF-beta1 through interference with Smad3 binding to the gene promoter.


Inflammatory and immune pathways in the pathogenesis of periodontal disease.
The pathogenesis of periodontitis involves a complex immune/inflammatory cascade that is initiated by the bacteria of the oral biofilm that forms naturally on the teeth. The susceptibility to periodontitis appears to be determined by the host response; specifically, the magnitude of the inflammatory response and the differential activation of immune pathways. The purpose of this review was to delineate our current knowledge of the host response in periodontitis. The role of innate immunity, the failure of acute inflammation to resolve (thus becoming chronic), the cytokine pathways that regulate the activation of acquired immunity and the cells and products of the immune system are considered. New information relating to regulation of both inflammation and the immune response will be reviewed in the context of susceptibility to, and perhaps control of, periodontitis.


Large-scale molecular dynamics simulations of bubble collapse in water: Effects of system size, water model, and nitrogen.
Molecular dynamics simulations in the microcanonical ensemble are performed to study the collapse of a bubble in liquid water using the single-site mW and the four-site TIP4P/2005 water models. To study system size effects, simulations for pure water systems are performed using periodically replicated simulation boxes with linear dimensions, L, ranging from 32 to 512 nm with the largest systems containing 8.7 × 106 and 4.5 × 109 molecules for the TIP4P/2005 and mW water models, respectively. The computationally more efficient mW water model allows us to reach converging behavior when the bubble dynamics results are plotted in reduced units, and the limiting behavior can be obtained through linear extrapolation in L-1. Qualitative differences are observed between simulations with the mW and TIP4P/2005 water models, but they can be explained by the models' differences in predicted viscosity and surface tension. Although bubble collapse occurs on time scales of only hundreds of picoseconds, the system sizes used here are sufficiently large to obtain bubble dynamics consistent with the Rayleigh-Plesset equation when using the models' thermophysical properties as input. For the conditions explored here, extreme heating of the interfacial water molecules near the time of collapse is observed for the larger mW water systems (but the model underpredicts the viscosity), whereas heating is less pronounced for the TIP4P/2005 water systems because its larger viscosity contribution slows the collapse dynamics. The presence of nitrogen within the bubble only starts to affect bubble dynamics near the very end of the initial collapse, leading to an incomplete collapse and strong rebound for the mW water model. Although nitrogen is non-condensable at 300 K, it becomes highly compressed and reaches a liquid-like density near the collapse point. We find that the dissolution of nitrogen is much slower than the movement of the collapsing water front, and the re-expansion of the dense nitrogen droplet gives rise to bubble rebound. The incompatibility of the collapse and dissolution time scales should be considered for continuum-scale modeling of bubble dynamics. We also confirm that the diffusion coefficient for dissolved nitrogen is insensitive to pressure as the liquid transitions from a compressed to a stretched state.


Coexisting malignant melanoma and vitiligo in a Nigerian.
A case of malignant melanoma associated with vitiligo in a middle aged Nigerian is presented. The Association has been well documented in some other parts of the world, but this is the first of its type in the West African subregion. The occurrence of vitiligo in melanoma patients is generally believed to be beneficial, and has been well recognised to presage long time survival for the patient. The course of events in our patient is still being observed.


Saccular aneurysm formation of the descending aorta associated with aortic coarctation in an infant.
Aneurysm of the descending aorta associated with CoA is an extremely rare congenital abnormality. In this report, we present a 16 months old female patient in whom cardiac catheterization had been performed which had revealed a segment of coarctation and saccular aneurysm in the descending aorta. The patient was operated and a 3x2 centimeters aneurysm which embraces the coarcted segment in descending aorta was resected. In summary, we present a case of saccular aortic aneurysm distal to aortic coarctation in an infant without any history of intervention or vascular inflammatory disease. Our case report seems to be the youngest patient in literature with this pathology.


Lactic acid bacteria efficiently protect human and animal intestinal epithelial and immune cells from enteric virus infection.
This study aimed to examine the potential antiviral activity of lactic acid bacteria (LAB) using animal and human intestinal and macrophage cell line models of non tumor origin. To this end, LAB strains selected on the basis of previous in vitro trials were co-incubated with cell line monolayers, which were subsequently challenged with rotavirus (RV) and transmissible gastroenteritis virus (TGEV). In order to elucidate the possible mechanism responsible for the antiviral activity, the induction of reactive oxygen species (ROS) release as well as the attachment ability of LAB on the cell lines was investigated. Various strains were found to exhibit moderate to complete monolayer protection against viral RV or TGEV disruption. Highest protection effects were recorded with the known probiotics Lactobacillus rhamnosus GG and Lactobacillus casei Shirota against both RV and TGEV, while notable antiviral activity was also attributed to Enterococcus faecium PCK38, Lactobacillus fermentum ACA-DC179, Lactobacillus pentosus PCA227 and Lactobacillus plantarum PCA236 and PCS22, depending on the cell line and virus combination used. A variable increase (of up to 50%) on the release of NO(-) and H(2)O(2) (ROS) was obtained when LAB strains were co-incubated with the cell lines, but the results were found to be LAB strain and cell line specific, apart from a small number of strains which were able to induce strong ROS release in more than one cell line. In contrast, the ability of the examined LAB strains to attach to the cell line monolayers was LAB strain but not cell line specific. Highest attachment ability was observed with L. plantarum ACA-DC 146, L. paracasei subsp. tolerans ACA-DC 4037 and E. faecium PCD71. Clear indications on the nature of the antiviral effect were evident only in the case of the L. casei Shirota against TGEV and with L. plantarum PCA236 against both RV and TGEV. In the rest of the cases, each interaction was LAB-cell line-virus specific, barring general conclusions. However, it is probable that more than one mechanism is involved in the antiviral effect described here. Further investigations are required to elucidate the underlying mode of action and to develop a cell line model as a system for selection of probiotic strains suited for farm animal applications.


Is disinvestment from alcohol and drug treatment services associated with treatment access, completions and related harm? An analysis of English expenditure and outcomes data.
The positive impact of substance use treatment is well-evidenced but there has been substantial disinvestment from publicly funded treatment services in England since 2013/2014. This paper examines whether this disinvestment from adult alcohol and drug treatment provision was associated with changes in treatment and health outcomes, including: treatment access, successful completions from treatment, alcohol-specific hospital admissions, alcohol-specific mortality and drug-related deaths. Annual administrative data from 2013/2014 to 2018/2019 was matched at local government level and multi-level time series analysis using linear mixed-effect modelling conducted for 151 upper-tier local authorities in England. Between 2013/2014 and 2018/2019, £212.2 million was disinvested from alcohol and drug treatment services, representing a 27% decrease. Concurrently, 11% fewer people accessed, and 21% fewer successfully completed, treatment. On average, controlling for other potential explanatory factors, a £10 000 disinvestment from alcohol and drug treatment services was associated with reductions in all treatment outcomes, including 0.3 fewer adults in treatment (95% confidence interval 0.16-0.45) and 0.21 fewer adults successfully completing treatment (95% % confidence interval 0.12-0.29). A £10 000 disinvestment from alcohol treatment was not significantly associated with changes in alcohol-specific hospital admissions or mortality, nor was disinvestment from drug treatment associated with the rate of drug-related deaths. Local authority spending cuts to alcohol and drug treatment services in England were associated with fewer people accessing and successfully completing alcohol and drug treatment but were not associated with changes in related hospital admissions and deaths.


Persistent clotting protein pathology in Long COVID/Post-Acute Sequelae of COVID-19 (PASC) is accompanied by increased levels of antiplasmin.
Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by acute clinical pathologies, including various coagulopathies that may be accompanied by hypercoagulation and platelet hyperactivation. Recently, a new COVID-19 phenotype has been noted in patients after they have ostensibly recovered from acute COVID-19 symptoms. This new syndrome is commonly termed Long COVID/Post-Acute Sequelae of COVID-19 (PASC). Here we refer to it as Long COVID/PASC. Lingering symptoms persist for as much as 6 months (or longer) after acute infection, where COVID-19 survivors complain of recurring fatigue or muscle weakness, being out of breath, sleep difficulties, and anxiety or depression. Given that blood clots can block microcapillaries and thereby inhibit oxygen exchange, we here investigate if the lingering symptoms that individuals with Long COVID/PASC manifest might be due to the presence of persistent circulating plasma microclots that are resistant to fibrinolysis. We use techniques including proteomics and fluorescence microscopy to study plasma samples from healthy individuals, individuals with Type 2 Diabetes Mellitus (T2DM), with acute COVID-19, and those with Long COVID/PASC symptoms. We show that plasma samples from Long COVID/PASC still contain large anomalous (amyloid) deposits (microclots). We also show that these microclots in both acute COVID-19 and Long COVID/PASC plasma samples are resistant to fibrinolysis (compared to plasma from controls and T2DM), even after trypsinisation. After a second trypsinization, the persistent pellet deposits (microclots) were solubilized. We detected various inflammatory molecules that are substantially increased in both the supernatant and trapped in the solubilized pellet deposits of acute COVID-19 and Long COVID/PASC, versus the equivalent volume of fully digested fluid of the control samples and T2DM. Of particular interest was a substantial increase in α(2)-antiplasmin (α2AP), various fibrinogen chains, as well as Serum Amyloid A (SAA) that were trapped in the solubilized fibrinolytic-resistant pellet deposits. Clotting pathologies in both acute COVID-19 infection and in Long COVID/PASC might benefit from following a regime of continued anticlotting therapy to support the fibrinolytic system function.


Functionality of mutations at conserved nucleotides in eukaryotic SECIS elements is determined by the identity of a single nonconserved nucleotide.
In eukaryotes, the specific cotranslational insertion of selenocysteine at UGA codons requires the presence of a secondary structural motif in the 3' untranslated region of the selenoprotein mRNA. This selenocysteine insertion sequence (SECIS) element is predicted to form a hairpin and contains three regions of sequence invariance that are thought to interact with a specific protein or proteins. Specificity of RNA-binding protein recognition of cognate RNAs is usually characterized by the ability of the protein to recognize and distinguish between a consensus binding site and sequences containing mutations to highly conserved positions in the consensus sequence. Using a functional assay for the ability of wild-type and mutant SECIS elements to direct cotranslational selenocysteine incorporation, we have investigated the relative contributions of individual invariant nucleotides to SECIS element function. We report the novel finding that, for this consensus RNA motif, mutations at the invariant nucleotides are tolerated to different degrees in different elements, depending on the identity of a single nonconserved nucleotide. Further, we demonstrate that the sequences adjacent to the minimal element, although not required for function, can affect function through their propensity to base pair. These findings shed light on the specific structure these conserved sequences may form within the element. This information is crucial to the design of strategies for the identification of SECIS-binding proteins, and hence the elucidation of the mechanism of selenocysteine incorporation in eukaryotes.


The scale of local adaptation in Mimulus guttatus: comparing life history races, ecotypes, and populations.
Fitness trade-offs between environments are central to the evolution of biodiversity. Although transplant studies often document fitness trade-offs consistent with local adaptation (LA), many have also found an advantage of foreign genotypes (foreign advantage (FA)). Understanding the mechanisms driving the magnitude and distribution of fitness variation requires comparative approaches that test the ecological scales at which these different patterns emerge. We used a common garden transplant experiment to compare the relative fitnesses of native vs foreign genotypes at three nested ecological scales within Mimulus guttatus: annual vs perennial life history races, perennial ecotypes across an elevational range, and populations within perennial elevational ecotypes. We integrated fitness across the life-cycle and decomposed LA vs FA into contributions from different fitness components. We found LA, measured as home-site advantage, between annual and perennial races and a trend towards LA among populations within montane habitats. Conversely, we found strong FA of low-elevation perennials in a montane environment. LA between life history races reflects the fitness advantages of adult survival and vegetative growth in a mesic environment. Within the perennial race, recent climate conditions or nonselective processes, such as dispersal limitation or mutational load, could explain FA of low-elevation perennials in a montane environment.


Multidrug Resistance of Gastric Cancer: The Mechanisms and Chinese Medicine Reversal Agents.
Chemotherapy is the main clinical treatment method of gastric cancer. Multidrug resistance (MDR) has become a common phenomenon with the development of tumors, which alleviates the effect of chemotherapy and makes it difficult to break the bottleneck of survival rate of advanced gastric cancer. Therefore, the exploration of MDR reversal agents for gastric cancer is the focus and also the difficulty of current treatment. Currently, the researches on the mechanisms of drug resistance in gastric cancer have been continuously deepened, which reveal different pathways and targets of MDR, laying a solid foundation for studying reversal agents. As a kind of natural medicine, traditional Chinese medicine (TCM) owns the characteristics of low toxicity, high safety and effectiveness. It can inhibit the occurrence, growth and metastasis of tumors, and reverse MDR via multiple pathways and mechanisms, the pathological function of which has become a research hotspot in recent years. TCM reversers are mainly divided into Chinese medicine monomers, Chinese patent medicines, and Chinese herbal compounds. With certain quantity and advantage, TCM reversers for MDR play an important role in the clinical treatment and show great potential in gastric cancer.


Effects of cyclic adenosine 3',5'-monophosphate on photoreceptor disc shedding and retinomotor movement. Inhibition of rod shedding and stimulation of cone elongation.
As a test of the hypothesis that cyclic nucleotides play a role in the regulation of retinomotor movements and disc shedding in the photoreceptor-pigment epithelial complex, we have used an in vitro eyecup preparation that sustains both disc shedding and cone retinomotor movements, Eyecups were prepared in white light from animals in which both shedding and cone movement had been blocked by 4 d of constant-light treatment. In eyecups incubated for 3 h in light, disc shedding was negligible and cones remained in the light-adapted (contracted) position. In eyecups incubated in darkness, however, a massive shedding response (dominated by rod photoreceptors) was induced, and at the same time cone photoreceptors elongated to their dark-adapted position. In eyecups incubated in light dbcAMP promoted cone elongation and thus mimicked darkness; the dbcAMP effect was potentiated by the phosphodiesterase inhibitors papaverine and 3-isobutylmethylxanthine. In eyecups incubated in darkness, on the other hand, both phosphodiesterase inhibitors and dbcAMP reduced the phagosome content of the pigment epithelium. The effects of dbcAMP on the cone elongation and rod shedding appear to be specific in that dbcGMP, adenosine, and adenosine 5'-monophosphate had no significant effect. Our results suggest that cAMP plays a role in the regulation of both retinomotor movements and disc shedding.


Association of hepatitis C virus replication complexes with microtubules and actin filaments is dependent on the interaction of NS3 and NS5A.
The hepatitis C virus (HCV) RNA replication complex (RC), which is composed of viral nonstructural (NS) proteins and host cellular proteins, replicates the viral RNA genome in association with intracellular membranes. Two viral NS proteins, NS3 and NS5A, are essential elements of the RC. Here, by using immunoprecipitation and fluorescence resonance energy transfer assays, we demonstrated that NS3 and NS5A interact with tubulin and actin. Furthermore, immunofluorescence microscopy and electron microscopy revealed that HCV RCs were aligned along microtubules and actin filaments in both HCV replicon cells and HCV-infected cells. In addition, the movement of RCs was inhibited when microtubules or actin filaments were depolymerized by colchicine and cytochalasin B, respectively. Based on our observations, we propose that microtubules and actin filaments provide the tracks for the movement of HCV RCs to other regions in the cell, and the molecular interactions between RCs and microtubules, or RCs and actin filaments, are mediated by NS3 and NS5A.


Change in self-reported health: A signal for early intervention in a medicare population.
Health plans and risk-bearing provider organizations seek information sources to inform proactive interventions for patients at risk of adverse health events. Interventions should take into account the strong relationship between social context and health. This retrospective cohort study of a Medicare Advantage population examined whether a change in self-reported health-related quality of life (HRQOL) signals a subsequent change in healthcare needs. A retrospective longitudinal analysis of administrative claims data was conducted for participants in a Medicare Advantage plan with prescription drug coverage (MAPD) who responded to 2 administrations of the Centers for Disease Control and Prevention 4-item Healthy Days survey within 6-18 months during 2015-2018. Changes in HRQOL, as measured by the Healthy Days instrument, were compared with changes in utilization and costs, which were considered to be a reflection of change in healthcare needs. A total of 48,841 individuals met inclusion criteria. Declining HRQOL was followed by increases in utilization and costs. An adjusted analysis showed that every additional unhealthy day reported one year after baseline was accompanied by an $8 increase in monthly healthcare costs in the subsequent six months for the average patient. Declining HRQOL signaled subsequent increases in healthcare needs and utilization. Findings suggest that HRQOL assessments in general, and the Healthy Days instrument in particular, could serve as a leading indicator of the need for interventions designed to mitigate poor health outcomes and rising healthcare costs. III.


Circulating TNF and mitochondrial DNA are major determinants of neutrophil phenotype in the advanced-age, frail elderly.
Tumor necrosis factor (TNF), a potent inflammatory cytokine, and mitochondrial DNA (mtDNA), a product of inflammation-induced tissue damage, increase with age ("inflammaging") and many chronic diseases. Peripheral blood neutrophils, a critical component of innate immunity, have also been shown to be altered with age, and are exceptionally sensitive to external stimuli. Herein, we describe that the phenotype of neutrophils from the advanced-age, frail elderly (ELD) is determined by levels of circulating TNF and mtDNA. Neutrophils from ELD donors are morphologically immature, and have higher levels of intracellular reactive oxygen species (ROS) and expression of the activation markers CD11b and HLA-DR. The frequency of CD11b(++) neutrophils correlated with plasma TNF, and recombinant TNF elevated neutrophil CD11b ex vivo and in vivo. Furthermore, neutrophils from aged TNF-deficient mice expressed CD11b similar to young counterparts. The frequency of HLA-DR(+) neutrophils, on the other hand, positively correlated with circulating mtDNA, which increased neutrophil HLA-DR expression in a dose-dependent manner ex vivo. Cell-surface TLR-9 expression, however, was unaltered on neutrophils from ELD donors. In summary, we provide novel evidence that products of age-related inflammation modulate neutrophil phenotype in vivo. Given this, anti-inflammatory therapies may prove beneficial in improving neutrophil functionality in the elderly.


Academy of breastfeeding medicine founder's lecture 2011: inequalities and inequities in breastfeeding: an international perspective.
Breastfeeding is the biological norm for infant feeding but is also a social construct. As such, its rates and practices are determined by the same social determinants that shape health inequalities and inequities. In the past 30 years, several reports have drawn attention to the changing pattern of breastfeeding inequalities across countries and population groups. Breastfeeding rates tend to fall and rise following a similar pattern everywhere, although at different times and speed. The role of women within families and societies, the routines of maternity hospitals and other healthcare services, and the pressure exerted by the baby food industry are among the factors that influence the time and speed of changes in breastfeeding rates and practices across countries and population groups. Inequities (i.e., inequalities considered unfair and avoidable by reasonable action) can be redressed by interventions for the protection, promotion, and support of breastfeeding. Evidence-based and quality-implemented support and promotion activities, if applied without an equity lens, may increase inequities. Activities for the protection of breastfeeding (e.g., implementation and enforcement of the International Code of Marketing of Breastmilk Substitutes; legislations, regulations, and policies to remove obstacles and barriers to good-quality breastfeeding support and to protect women and mothers in the workforce; elimination of obstacles and barriers to breastfeeding anywhere, anyhow, and anytime mothers want) apply to all women and are less dependent on take up by the target population. If well designed and enforced, protective interventions contribute to reducing inequalities and inequities and to delivering promotion and support activities more effectively.


Selective coronary arteriography.
The technique of selective coronary arteriography, as described originally by Sones, was employed in 255 patients. Successful catheterization of both coronary arteries was carried out in 88% of these patients, and in the last 100 examinations both coronary arteries were entered in 95 patients. Selective coronary arteriography is a useful diagnostic tool but is a potentially hazardous form of examination as we encountered four episodes of ventricular fibrillation in the present series.


Use of non-formulary high-cost medicines in an Australian public hospital.
Background Clinicians prescribe high-cost medicines for rare diseases and nonapproved indications when conventional therapies have failed. Objective To examine the use of non-formulary high-cost medicines at an Australian public hospital. Methods Retrospective audit of individual patient use applications for nonformulary medicines costing more than $5000 AUD per year at a large tertiary referral hospital in Adelaide, South Australia over a 12-month study period from January 2015 to December 2015. Main outcome measures Total cost of non-formulary high-cost medicines, medication class, indications for use, level of supporting evidence and proposed monitoring outcomes. Results Eighty-seven individual patient use applications were examined. All except one were approved, at a total cost of $1,339,203 AUD. The most common drug classes were anti-CD20 (n = 33, 38%), combined antiretrovirals (n = 10, 11%) and TNF-alpha antagonists (n = 10, 11%). There were 56 indications for these medicines with the majority being inflammatory conditions (n = 52, 60%), followed by infections (n = 14, 16%) and malignancies (n = 14, 16%). Of the first-time individual patient use applications (n = 63), there were 25 applications (40%) that provided a case series as supporting evidence. Approximately half of new individual patient use applications (n = 32) proposed an objective monitoring outcome, but few (n = 13, 21%) contained sufficient information to allow a third party to determine efficacy of the medication. Conclusions Non-formulary high-cost medicines are being used for a broad range of indications based largely on low levels of evidence. Prospective definition of an adequate response to treatment and reporting of these outcomes is required to improve the evidence-base and to aid decision-making for subsequent treatment courses.


Soluble epoxide hydrolase inhibition improves coronary endothelial function and prevents the development of cardiac alterations in obese insulin-resistant mice.
This study addressed the hypothesis that inhibiting the soluble epoxide hydrolase (sEH)-mediated degradation of epoxy-fatty acids, notably epoxyeicosatrienoic acids, has an additional impact against cardiovascular damage in insulin resistance, beyond its previously demonstrated beneficial effect on glucose homeostasis. The cardiovascular and metabolic effects of the sEH inhibitor trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid (t-AUCB; 10 mg/l in drinking water) were compared with those of the sulfonylurea glibenclamide (80 mg/l), both administered for 8 wk in FVB mice subjected to a high-fat diet (HFD; 60% fat) for 16 wk. Mice on control chow diet (10% fat) and nontreated HFD mice served as controls. Glibenclamide and t-AUCB similarly prevented the increased fasting glycemia in HFD mice, but only t-AUCB improved glucose tolerance and decreased gluconeogenesis, without modifying weight gain. Moreover, t-AUCB reduced adipose tissue inflammation, plasma free fatty acids, and LDL cholesterol and prevented hepatic steatosis. Furthermore, only the sEH inhibitor improved endothelium-dependent relaxations to acetylcholine, assessed by myography in isolated coronary arteries. This improvement was related to a restoration of epoxyeicosatrienoic acid and nitric oxide pathways, as shown by the increased inhibitory effects of the nitric oxide synthase and cytochrome P-450 epoxygenase inhibitors l-NA and MSPPOH on these relaxations. Moreover, t-AUCB decreased cardiac hypertrophy, fibrosis, and inflammation and improved diastolic function, as demonstrated by the increased E/A ratio (echocardiography) and decreased slope of the end-diastolic pressure-volume relation (invasive hemodynamics). These results demonstrate that sEH inhibition improves coronary endothelial function and prevents cardiac remodeling and diastolic dysfunction in obese insulin-resistant mice.


Characterization of a monoclonal antibody reacting with the free human luteinizing hormone beta-subunit.
Measurement of serum luteinizing hormone (hLH) is important for the detection and follow-up of patients with pathological processes of the reproductive axis. Detection of the uncombined form of the beta-subunit of hLH, or free hLH beta, has proved to be of clinical interest in the recognition of gonadotroph adenomas. As no monoclonal antibody specific for the free hLH beta is at present available, we elicited monoclonal antibodies using free hLH beta as an immunogen. An antibody, named BLH01, was selected for its specific binding to the free beta-subunit, and its antibody-binding site was characterized at the molecular level, emphasizing the importance of amino acid residues located between the disulfide-bonded Cys93 and Cys100. This region has been demonstrated as being particularly critical for the specific binding of lutropic hormones to their receptors. Topographic assignment of the epitope recognized by BLH01 was then achieved by cross-matching studies based on a library of antibodies directed to hLH beta, and the location of some epitopes on the three-dimensional model of the beta-subunit is proposed. A two-site immunoassay based on BLH01 as capture antibody was then developed. This assay, using BLH01, may constitute a simple, sensitive and highly specific procedure for assessing the clinical usefulness of measuring free hLH beta, particularly for the diagnosis and follow-up of patients presenting with pituitary adenomas.


Direct acting antiviral HCV treatment does not influence renal function.
HCV infection is associated with chronic kidney disease due to several mechanisms. Patients treated with interferon-based regimens demonstrate improved renal function and reduced incidence of chronic kidney disease. There is scarce evidence on the effect of direct acting antiviral regimens (DAAs) on renal function.We evaluated serial measures of renal function in a cohort of HCV-infected participants following completion of DAA-based treatment regimens.Measures of glomerular filtration rate (GFR) were estimated by the CKD-EPI equation. Data was recorded at end of treatment, and at 6-12 months, 12-24 months, and greater than 24 months following treatment completion. Group-based trajectory modeling was used to determine distinct GFR trajectories. Predictors of group membership were determined by multinomial regression analysis.Six trajectories were identified. One trajectory comprising 27% of the cohort demonstrated declining renal function and the others demonstrated no change in renal function over time. Baseline GFR did not predict SVR. Diabetes was associated with lower post-treatment GFR but patients with diabetes did not demonstrate a decrease in GFR over the period of evaluation. Cirrhosis and SVR were not significant predictors of GFR or GFR trajectory.There is no clinically relevant change in renal function among the majority of HCV-infected patients following completion of DAA-based treatments. Renal function does not influence the efficacy of DAA-based regimens. No consistent effect of DAA treatment and/or SVR on renal function was observed over a 2-year period following treatment completion.


A discussion on implementing pooling detection tests of novel coronavirus (SARS-CoV-2) for a large population.
A pooled sample analysis strategy for novel coronavirus (severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2)) is proposed for a large population in this paper. The population to be tested is divided into divisions based on earlier observed detection rate of SARS-CoV-2 first. Samples collected are then grouped in appropriate pooled size. The number of tests per person in that population is expressed as a function of two variables: the observed detection rate and the pooled size or number of samples grouped. The minimum number of tests per person can be further shown to be a function of only one of these two variables, because these two parameters are found to be related at this minimum. A management scheme on grouping the samples is proposed in order to reduce the number of tests, to save time, which is of utmost importance in fighting an epidemic. The proposed testing scheme will be useful for supporting the government in making decisions to handle regular routine detection tests for identifying asymptomatic patients and implementing health code system in large population of millions of citizens. Another important point is to use smaller number of test kits, allowing more resources to speed up the mass screening tests, particularly in places not so rich.


Programs Addressing Food Security for First Nations Peoples: A Scoping Review.
Access to food is a right that every individual must have to ensure a standard of living that is sufficient for maintaining good health and wellbeing. This review, developed and implemented by a team of First Nations and non-First Nations peoples, aimed to scope the literature on programs addressing food security for First Nations peoples in Australia, Aotearoa/New Zealand, Canada, and the United States of America. Collectively, First Nations groups share continued traumas, disadvantages, and devastation brought upon them as a result of British colonisation. Despite the impacts of colonial conquest, the resilience of First Nations peoples continues through the fight for self-determination, sovereignty, equity, and equality. Three databases and grey literature were searched from 2010. Two reviewers completed screening, data extraction, and critical appraisal. Nine food security programs were included in this review. Five were from the United States of America and four from Canada, with no program from Australia or Aotearoa/New Zealand meeting the inclusion criteria. The programs that appear to be most suitable for addressing food security for First Nations peoples were participatory in design, had community governance, integrated cultural knowledge and food systems to increase the accessibility and availability of cultural foods, incorporated educational components, and utilized collaborations among various agencies. Findings showed that while it is important to address short-term emergency food relief, the aim should be sustainable food security through a longer-term system and policy change underpinned by co-designed research and evaluation.


Skin surveillance intentions among family members of patients with melanoma.
First-degree relatives of individuals diagnosed with melanoma are at increased disease risk. However, many first-degree relatives do not receive a periodic total cutaneous examination from a health care provider or engage in regular skin self-examination. The goal of this study was to identify correlates of total cutaneous examination and skin self-examination intentions among first-degree relatives of melanoma patients, thus providing insight on factors that should be targeted in future intervention research. The participants were 545 first-degree relatives of melanoma patients at increased disease risk due to their risk factor profile and lack of skin surveillance behaviors. Participants completed a telephone survey regarding their total cutaneous examination and skin self-examination intentions and potential correlates, including demographics, medical factors, psychological factors, knowledge, and social influence factors. Intentions to receive a total cutaneous examination were higher among first-degree relatives with more education, those perceiving higher benefits and lower barriers to an examination, and those reporting greater physician and family support. Intentions to receive a skin self-examination were higher among those with higher benefits and lower barriers to self-examination, and higher family support. Interventions to promote skin surveillance behaviors among first-degree relatives of melanoma patients should highlight the benefits of early detection of melanoma, address barriers to receipt of total cutaneous examination and engagement in skin self-examination, and promote support from physicians and family members.


Pulmonary Embolism in COVID-19 and the Unanswered Questions.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has ravaged the global society as we know it. As almost a dozen pharmaceutical agents go into randomized controlled clinical trials, multiple studies have surfaced trying to associate a hypercoagulable state to coronavirus disease 2019 (COVID-19) patients. We report two COVID-19 cases who presented with occlusive pulmonary embolism (PE) strongly supporting a hypercoagulable state incurred by SARS-CoV-2. This is significant as it is one of the early reports of such an initial presentation of COVID-19 in the USA. Through our report, we invite the medical community to share a perspective about long-term management guidelines for SARS-CoV-2 associated venous thromboembolism (VTE) and prompt future research.


Temperature Impact in LoRaWAN-A Case Study in Northern Sweden.
LoRaWAN has become popular as an IoT enabler. The low cost, ease of installation and the capacity of fine-tuning the parameters make this network a suitable candidate for the deployment of smart cities. In northern Sweden, in the smart region of Skellefteå, we have deployed a LoRaWAN to enable IoT applications to assist the lives of citizens. As Skellefteå has a subarctic climate, we investigate how the extreme changes in the weather happening during a year affect a real LoRaWAN deployment in terms of SNR, RSSI and the use of SF when ADR is enabled. Additionally, we evaluate two propagation models (Okumura-Hata and ITM) and verify if any of those models fit the measurements obtained from our real-life network. Our results regarding the weather impact show that cold weather improves the SNR while warm weather makes the sensors select lower SFs, to minimize the time-on-air. Regarding the tested propagation models, Okumura-Hata has the best fit to our data, while ITM tends to overestimate the RSSI values.


Object relations assessment predicts outpatients' attrition, early working alliance and psychotherapy process.
This study investigated the predictive utility of assessing clients' object relations functioning to prognosticate therapy dropout, quality of the early working alliance and psychotherapy process events reported by clients after their first five sessions. Clients accepting a recruitment invitation were administered the thematic apperception test (TAT) shortly after intake, and those still in treatment three to four sessions later rated the working alliance and psychotherapy process events. Participants were 47 clients beginning psychotherapy with advanced doctoral practicum students at a university-based community-serving training clinic. The Social Cognition and Object Relations Scales (SCORS), an object relations scoring system for TAT stories, was used to assess object relations functioning. The California Psychotherapy Alliance Scales-Patient form measured four dimensions of the alliance. The Therapy Orientation Process Scales, created from the Psychotherapy Process Q-set, measured clients' perceptions of therapy process events as involving relatively more psychodynamic or cognitive behavioural techniques. The SCORS significantly predicted remaining in therapy, client ratings of stronger working alliance and typical therapy process as more psychodynamic than cognitive behavioural. SCORS complexity of representations and capacity for emotional investment in relationships scales were the strongest predictors, especially of the patient commitment facet of the alliance. The findings suggest that (1) beginning psychotherapy with object relations assessment can be useful for treatment planning and (2) the relationship between clients' object relations functioning, working alliance and greater sensitivity to psychodynamic than to cognitive behavioural interventions depends on the aspect of object relations that is being assessed.


Sensorimotor-independent prefrontal activity during response inhibition.
A network of brain regions involving the ventral inferior frontal gyrus/anterior insula (vIFG/AI), presupplementary motor area (pre-SMA) and basal ganglia has been implicated in stopping impulsive, unwanted responses. However, whether this network plays an equal role in response inhibition under different sensorimotor contexts has not been tested systematically. Here, we conducted an fMRI experiment using the stop signal task, a sensorimotor task requiring occasional withholding of the planned response upon the presentation of a stop signal. We manipulated both the sensory modality of the stop signal (visual versus auditory) and the motor response modality (hand versus eye). Results showed that the vIFG/AI and the preSMA along with the right middle frontal gyrus were commonly activated in response inhibition across the various sensorimotor conditions. Our findings provide direct evidence for a common role of these frontal areas, but not striatal areas in response inhibition independent of the sensorimotor contexts. Nevertheless, these three frontal regions exhibited different activation patterns during successful and unsuccessful stopping. Together with the existing evidence, we suggest that the vIFG/AI is involved in the early stages of stopping such as triggering the stop process while the preSMA may play a role in regulating other cortical and subcortical regions involved in stopping.


Blood pressure regulation V: in vivo mechanical properties of precapillary vessels as affected by long-term pressure loading and unloading.
Recent studies are reviewed, concerning the in vivo wall stiffness of arteries and arterioles in healthy humans, and how these properties adapt to iterative increments or sustained reductions in local intravascular pressure. A novel technique was used, by which arterial and arteriolar stiffness was determined as changes in arterial diameter and flow, respectively, during graded increments in distending pressure in the blood vessels of an arm or a leg. Pressure-induced increases in diameter and flow were smaller in the lower leg than in the arm, indicating greater stiffness in the arteries/arterioles of the leg. A 5-week period of intermittent intravascular pressure elevations in one arm reduced pressure distension and pressure-induced flow in the brachial artery by about 50%. Conversely, prolonged reduction of arterial/arteriolar pressure in the lower body by 5 weeks of sustained horizontal bedrest, induced threefold increases of the pressure-distension and pressure-flow responses in a tibial artery. Thus, the wall stiffness of arteries and arterioles are plastic properties that readily adapt to changes in the prevailing local intravascular pressure. The discussion concerns mechanisms underlying changes in local arterial/arteriolar stiffness as well as whether stiffness is altered by changes in myogenic tone and/or wall structure. As regards implications, regulation of local arterial/arteriolar stiffness may facilitate control of arterial pressure in erect posture and conditions of exaggerated intravascular pressure gradients. That increased intravascular pressure leads to increased arteriolar wall stiffness also supports the notion that local pressure loading may constitute a prime mover in the development of vascular changes in hypertension.


13-cis-retinoic acid plus interferon alpha-2a: highly active systemic therapy for squamous cell carcinoma of the cervix.
Chemotherapeutic study of cervical squamous cell carcinoma has shown some positive results. Complete plus partial (overall) response rates of 15%-35% (complete response rate, less than 5%) were achieved with the use of a small number of cytotoxic single agents in patients with advanced disease. In addition, overall response rates of 60%-70% (complete response rates, 10%-20%) were achieved with cisplatin-based, multiagent regimens in patients with primary, locally advanced disease. However, the lack of clear evidence that existing chemotherapy can achieve a survival benefit, coupled with the worldwide annual deaths of hundreds of thousands of women from cervical cancer, indicates the urgent need for effective systemic therapy for this disease. In view of the preclinical and clinical evidence that supports testing of the novel combination of 13-cis-retinoic acid (13-cRA) plus interferon-alpha (IFN-alpha) in cervical squamous cell carcinoma, we conducted a phase II study of this regimen in locally advanced disease. Twenty-six patients with untreated, locally advanced squamous cell carcinoma of the cervix were treated daily for at least 2 months with oral 13-cRA (1 mg/kg) and subcutaneous recombinant human IFN alpha-2a (6 million units). In 21 patients (81%), the disease was stage II or higher. Thirteen patients (50%) experienced major responses (tumor regression greater than or equal to 50%) in association with resolution of symptoms; one achieved complete response, and 12 experienced partial response. Seven with partial response are improving further, four are being maintained in partial response, and one responder has relapsed during therapy. The response rate is 58% (11 of 19) in patients with stage IIB or higher disease and 66% (10 of 15) in patients with bulky disease (at least one dimension greater than or equal to 10 cm). Of the 13 non-responders, nine have stable disease and four have had disease progression during therapy. Toxicity was minimal. These preliminary results indicate that systemic 13-cRA plus IFN alpha-2a is a highly active, well-tolerated therapy for locally advanced cervical cancer.


Preferences and attitudes toward approaches to depression relapse/recurrence prevention among pregnant women.
Patient preferences are increasingly recognized as important in clinical research and the delivery of evidence based practice in psychology. Although the prevention of depressive relapse/recurrence among perinatal women is an important public health goal, little is known about pregnant women's preferences and attitudes toward relapse/recurrence prevention interventions. Such information is important given low rates of care seeking among this population, and the potential for a relapse/recurrence prevention to avert negative outcomes among both vulnerable women and their offspring. Pregnant women seeking routine prenatal care in obstetric clinics (n = 200) were surveyed to assess their preferences for and attitudes about psychotherapy and pharmacological approaches to relapse/recurrence prevention. Women preferred psychotherapy (mindfulness based cognitive therapy and interpersonal therapy) more so than pharmacotherapy and reported significantly more favorable perceptions of the psychotherapy as compared to pharmacotherapy approaches to depression relapse/recurrence prevention. Results suggest also that depression history is important to consider in evaluating women's preferences and attitudes. Clinical and research implications of these findings are discussed.


[Application of real-time fluorescence quantitative polymerase chain reaction techniques for prenatal diagnosis of Down syndrome].
To investigate the value of real-time fluorescence quantitative PCR in diagnosis of Down syndrome with uncultured amniotic cells. The uncultured amniocytes of 80 fetuses who were confirmed disomy 21 by chromosome analysis and 5 fetuses detected trisomy 21 and peripheral blood samples of 7 children diagnosed as Down syndrome were collected to extract gDNA and the real-time fluorescence quantitative PCR method was used to detect the original copies of Down syndrome critical region gene(3) (DSCR(3)) and GAPDH gene and then the ratio of DSCR(3)/GAPDH was calculated. The PCR product ratios of DSCR(3) to GAPDH in trisomy 21 from amniocytes and peripheral blood were ranged from 1.64 to 1.98 while in the normal control the ratio was only from 0.46 to 1.30. Real-time fluorescence quantitative PCR is a valuable method for rapid and accurate prenatal diagnosis of Down syndrome in uncultured amniotic cells.


Cell surface antigens of human bladder cancer defined by mouse monoclonal antibodies.
Mouse monoclonal antibodies were obtained by immunization with cultured human bladder cancer or lysates of bladder papilloma. They identify 11 distinct antigenic systems as defined by serological analysis of cultured cells and studies of antigen distribution in normal and neoplastic tissues. The most restricted of these antigens, Om5, defines a subset of bladder tumors. Om5 is not detected in normal bladder urothelium or in any other normal or malignant tissue. T101 and JP165 are also subset markers for bladder cancer that are not detected in normal tissues. T16, T43, T87, and J143 (antigens represented on many cultured cells) are found in specific areas of the normal urinary tract and in a distinctive range of other normal and malignant cell types--e.g., T16 expression in pluristratified epithelium of skin, exocervix, and esophagus. T138 antigen is also a common feature of cultured cell lines, but its expression in sections of normal tissues is restricted to endothelial cells. In contrast, T110 is poorly represented on cultured cells but can be detected in culture supernatants. Localization of T110 in normal tissues showed that it is a component of the extracellular matrix. All determinants detected by this series of antibodies are heat labile and not related to A, B, H, I, Lewis blood group antigens. Six of the antibodies immunoprecipitated glycoproteins from radiolabeled cell lysates: AbT16 (Mrs 48,000 and 42,000), AbT87 (Mr 60,000), AbT43 (Mr 85,000), AbJ143 (Mr 140,000, 120,000, 60,000), AbT43 (Mr 85,000), AbJ143 (Mr 140,000, 120,000, and 30,000), and AbT110 (Mr 240,000).


Metabolic fingerprinting of cell types in mouse skeletal muscle by combining TOF-SIMS with immunofluorescence staining.
Skeletal muscle tissue is composed of various muscle cell types which differ in physiological functions. Changes in cell type composition of skeletal muscle are associated with the development of metabolic diseases. Skeletal muscle cell types are currently distinguished by immunofluorescence (IF) staining based on myosin heavy chain (MHC) isoform difference. However, it remains a challenge to provide metabolic fingerprints of different muscle cell types by IF staining. Therefore, in this study, we proposed a method to examine metabolite distribution within different cell types by time-of-flight secondary ion mass spectrometry (TOF-SIMS) with high spatial resolution. Skeletal muscle samples from C57/BL6 mice were obtained by slicing. Cell types in TOF-SIMS images were labelled corresponding to IF images from the same region of serially cut sections. Mass spectra corresponding to individual muscle cells were extracted to compare metabolic fingerprints among cell types. Skeletal muscle cells were classified into two clusters based on the mass spectra of individual cells. Unsaturated diacylglycerol (DG) and fatty acid (FA) species were found to be distributed in a cell-type dependent manner. Moreover, relative quantification showed that the content of unsaturated DGs, oleic acid and linoleic acid was higher in type I and type IIA cells than in type IIB cells. TOF-SIMS in combination with IF enables us to directly visualize metabolite distribution in different cell types, to find potential biomarkers for cell type classification. TOF-SIMS imaging coupled with IF staining has been proved to be a promising tool for metabolic fingerprinting of different skeletal muscle cell types.


Amlodipine and calcineurin inhibitor-induced nephrotoxicity following allogeneic hematopoietic stem cell transplant.
Studies in the renal transplant population have suggested calcium-channel blockers (CCBs) may protect against calcineurin inhibitor (CNI)-induced nephrotoxicity. However, this has not been evaluated in the hematopoietic stem cell transplant (HSCT) population. This retrospective study reviews data from 350 consecutive patients who underwent allogeneic HSCT to determine whether amlodipine improved renal outcomes. Subject data included up to one year from CNI initiation. Patients in the amlodipine group (n = 130) received an average of 143 days treatment with amlodipine and experienced a smaller decrease in creatinine clearance (CrCl) through day 180. At day 30, change in CrCl was -17.4 mL/min in the amlodipine cohort and -33.8 mL/min in the control (P < 0.001). At day 180, change in CrCl was -40.9 and -50.6 mL/min, respectively (P = 0.005). Incidence of hospitalization with acute kidney injury (AKI) was significantly lower in patients receiving amlodipine, 7.7% (10/132) vs 16.4% (36/220) (hazard ratio [HR] 0.44; 95% confidence interval [CI] 0.22-0.89). Median blood pressure in the amlodipine group remained <132/78 through day 360. Our data support the use of amlodipine for hypertension in the allogeneic HSCT population and provide evidence suggesting that CCBs protect against CNI-induced nephrotoxicity.


Distribution and development of nicotinic acetylcholine receptor subtypes in the optic tectum of Rana pipiens.
Acetylcholine allows the elicitation of visually evoked behaviors mediated by the frog optic tectum, but the mechanisms behind its effects are unknown. Although nicotinic acetylcholine receptors (nAChRs) exist in the tectum, their subtype has not been assessed. By using quantitative autoradiography, we examined the binding of [(3)H]cytisine and [(125)I]alpha-bungarotoxin in the laminated tectum. In mammalian systems, these radioligands bind with high affinity to alpha4 nAChR subunits and alpha7 nAChR subunits, respectively. [(3)H]Cytisine demonstrated high specific binding in adult frogs in retinorecipient layer 9, intermediate densities in layer 8, and low binding in layers 1-7 of the tectum. [(3)H]Cytisine binding was significantly higher in the tecta of adults than in those of tadpoles. Lesioning the optic nerve for 6 weeks decreased [(3)H]cytisine binding in layers 8/9 by 70+/-1%, whereas 6-month lesions decreased binding by 76+/-3%. Specific binding of [(125)I]alpha-bungarotoxin in adults was present only at intermediate levels in tectal layers 8 and 9, and undetectable in the deeper tectal layers. However, the nucleus isthmi, a midbrain structure reciprocally connected to the tectum, exhibited high levels of binding. There were no significant differences in tectal [(125)I]alpha-bungarotoxin binding between tadpoles and adults. Six-week lesions of the optic nerve decreased tectal [(125)I]alpha-bungarotoxin binding by 33+/-10%, but 6-month lesions had no effect. The pharmacokinetic characteristics of [(3)H]cytisine and [(125)I]alpha-bungarotoxin binding in the frog brain were similar to those demonstrated in several mammalian species. These results indicate that [(3)H]cytisine and [(125)I]alpha-bungarotoxin identify distinct nAChR subtypes in the tectum that likely contain non-alpha7 and alpha7 subunits, respectively. The majority of non-alpha7 receptors are likely associated with retinal ganglion cell terminals, whereas alpha7-containing receptors appear to have a different localization.


Polymorphisms in NAT2, CYP2D6, CYP2C19 and GSTP1 and their association with prostate cancer.
The development of prostate cancer is dependent on heredity, androgenic influences, and exposure to environmental agents. A high intake of dietary fat is associated with an increased risk of prostate cancer, either through influence on steroid hormone profiles or through production of carcinogenic compounds that require biotransformation by enzymes. The polymorphic glutathione S-transferase (GST), N-acetyltransferase (NAT), and cytochrome P450 (CYP) enzymes are of particular interest in prostate cancer susceptibility because of their ability to metabolize both endogenous and exogenous compounds, including dietary constituents. Association between different NAT2, CYP2D6, CYP2C19 and GSTP1 genotypes and prostate cancer was studied in a Swedish and Danish case-control study comprising 850 individuals. The combined Swedish and Danish study population was analysed by polymerase chain reaction for the NAT2 alleles *4, *5A, *5B, *5C, *6 and *7, and for the CYP2D6 alleles *l, *3 and *4. The Swedish subjects were also analysed for the CYP2C19 alleles *1 and *2, and the GSTP1 alleles *A, *B and *C. No association was found between prostate cancer and polymorphisms in NAT2, CYP2D6, CYP2C19 or GSTP1. An association between CYP2D6 poor metabolism and prostate cancer was seen among smoking Danes; odds ratio 3.10 (95% confidence interval 1.07; 8.93), P = 0.03, but not among smoking Swedes; odds ratio 1.19 (95% confidence interval 0.41; 3.42), P = 0.75. Smoking is not a known risk factor for prostate cancer, and the association between CYP2D6 poor metabolism and prostate cancer in Danish smokers may have arisen by chance.


Fatigue and sleep-disordered breathing in multiple sclerosis: a clinically relevant association?
Background. Fatigue in patients with multiple sclerosis (MS) is highly prevalent and severely impacts quality of life. Recent studies suggested that sleep-disordered breathing (SDB) significantly contributes to fatigue in MS. Study Objective. To evaluate the importance of routine respirography in MS patients with severe fatigue and to explore the effects of treatment with continuous positive airway pressure (CPAP). Patients and Methods. We prospectively assessed the presence of severe fatigue, as defined by a score of ≥5.0 on the Fatigue Severity Scale (FSS), in 258 consecutive MS patients. Ninety-seven patients (38%) suffered from severe fatigue, whereof 69 underwent overnight respirography. Results. We diagnosed SDB in 28 patients (41%). Male sex was the only independent associate of SDB severity (P = 0.003). CPAP therapy in 6 patients was associated with a significant reduction of FSS scores (5.8 ± 0.5 versus 4.8 ± 0.6, P = 0.04), but the scores remained pathological (≥4.0) in all patients. Conclusion. Respirography in MS patients with severe fatigue should be considered in daily medical practice, because SDB frequency is high and CPAP therapy reduces fatigue severity. However, future work is needed to understand the real impact of CPAP therapy on quality of life in this patient group.


Making sense of electrical sense in crayfish.
The five sensory modalities of humans are also found in a wide range of invertebrates. Other vertebrates have evolved additional special senses, such as the magnetic sense, which are also found in some invertebrates. However, there remain a few sensory abilities that curiously appear to be found in either vertebrates or invertebrates, but not both. For example, electrosensitivity - the ability to detect electric fields in water - which should benefit vertebrates and invertebrates alike, is apparently only used by vertebrates. However, recent reports suggest that some invertebrates could have an electric sense. Here we examine that possibility further and demonstrate a behavioural threshold to low-level electrical fields in two freshwater invertebrates. The responses are not low enough for them to detect the Earth's magnetic field as some other electroreceptive species can do, but sufficiently low for them to use in navigation or prey and predator detection. This finding challenges the current view of the sensory world of aquatic invertebrates and has implications for the evolution of this ability.


[Elemental status of the medical personnel of the emergency medical services in the city of Khanty-Mansiysk].
Spectrometric analysis of hair from 110 medical workers (54--from the Emergency medical services and 56--from polyclinics) was performed with the use of atomic emission spectrometry and mass spectrometry, inductively coupled argon plasma spectrometry (AES-ISP) methods. There were revealed features of the elemental status of the medical personnel of the Emergency medical services: a deficiency of Mg, K and Li was typical for this group (presented more then in half of cases).


Megalin (gp330): a putative endocytic receptor for thyroglobulin (Tg).
Megalin (gp330) is a large glycoprotein receptor found mainly on a group of absorptive epithelial cells, including renal proximal tubule, epididymal and thyroid cells. Megalin has been shown to bind multiple, unrelated ligands, mainly in vitro, and to mediate endocytosis of ligandsin cultured cells. However, physiologic ligands of megalin are largely unknown. In the present study we have demonstrated that purified rat megalin binds rat thyroglobulin (Tg) in solid phase assays, with anestimated Kd of 9.2+/-0.6 nM. Binding was calcium dependent and was almost completely inhibited by excess Tg, by three megalin ligands - lactoferrin, lipoprotein lipase and apolipoprotein J- and by the receptor associated protein (RAP), which inhibits binding of all megalin ligands. Three anti-megalin antibodies partially inhibited Tg binding to megalin. 125I labeled Tg bound to megalin was released by EDTA and heparin; the released product was shown by SDS-PAGE and autoradiography to be 660 kD (dimeric) Tg. However, an immunoblotting experiment showed binding of megalin both to monomeric (330 kD) and dimeric Tg. We propose that megalin, which is known to mediate ligand endocytosis and is found on the apical surface of thyrocytes, may participate in the endocytosis of Tg from the colloid, a process that is required for hormone release from Tg.


The FOX transcription factor Hcm1 regulates oxidative metabolism in response to early nutrient limitation in yeast. Role of Snf1 and Tor1/Sch9 kinases.
Within Saccharomyces cerevisiae, Hcm1is a member of the forkhead transcription factor family with a role in chromosome organization. Our group recently described its involvement in mitochondrial biogenesis and stress resistance, and reports here that Hcm1 played a role in adaptation to respiratory metabolism when glucose or nitrogen was decreased. Regulation of Hcm1 activity occurs in at least three ways: i) protein quantity, ii) subcellular localization, and iii) transcriptional activity. Transcriptional activity was measured using a reporter gene fused to a promoter that contains a binding site for Hcm1. We also analyzed the levels of several genes whose expression is known to be regulated by Hcm1 levels and the role of the main kinases known to respond to nutrients. Lack of sucrose-nonfermenting (Snf1) kinase increases cytoplasmic localization of Hcm1, whereas Δtor1 cells showed a mild increase in nuclear Hcm1. In vitro experiments showed that Snf1 clearly phosphorylates Hcm1 while Sch9 exerts a milder phosphorylation. Although in vitroTor1 does not directly phosphorylate Hcm1, in vivo rapamycin treatment increases nuclear Hcm1. We conclude that Hcm1 participates in the adaptation of cells from fermentation to respiratory metabolism during nutrient scarcity. According to our hypothesis, when nutrient levels decrease, Snf1 phosphorylates Hcm1. This results in a shift from the cytoplasm to the nucleus and increased transcriptional activity of genes involved in respiration, use of alternative energy sources, NAD synthesis and oxidative stress resistance.


Capturing the Flexibility of a Protein-Ligand Complex: Binding Free Energies from Different Enhanced Sampling Techniques.
Enhanced sampling techniques are a promising approach to obtain reliable binding free-energy profiles for flexible protein-ligand complexes from molecular dynamics (MD) simulations. To put four popular enhanced sampling techniques to a biologically relevant and challenging test, we studied the partial dissociation of an antigenic peptide from the Major Histocompatibility Complex I (MHC I) HLA-B*35:01 to systematically investigate the performance of umbrella sampling (US), replica exchange with solute tempering 2 (REST2), bias exchange umbrella sampling (BEUS, or replica-exchange umbrella sampling), and well-tempered metadynamics (MTD). With regard to the speed of sampling and convergence, the peptide-MHC I complex (pMHC I) under study showcases intrinsic strengths and weaknesses of the four enhanced sampling techniques used. We found that BEUS can best handle the sampling challenges that arise from the coexistence of an enthalpically and an entropically stabilized free-energy minimum in the pMHC I under study. These findings might also be relevant for other flexible biomolecular systems with competing enthalpically and entropically stabilized minima.