Simulation of FRET dyes allows quantitative comparison against experimental data.
Fully understanding biomolecular function requires detailed insight into the systems' structural dynamics. Powerful experimental techniques such as single molecule Förster Resonance Energy Transfer (FRET) provide access to such dynamic information yet have to be carefully interpreted. Molecular simulations can complement these experiments but typically face limits in accessing slow time scales and large or unstructured systems. Here, we introduce a coarse-grained simulation technique that tackles these challenges. While requiring only few parameters, we maintain full protein flexibility and include all heavy atoms of proteins, linkers, and dyes. We are able to sufficiently reduce computational demands to simulate large or heterogeneous structural dynamics and ensembles on slow time scales found in, e.g., protein folding. The simulations allow for calculating FRET efficiencies which quantitatively agree with experimentally determined values. By providing atomically resolved trajectories, this work supports the planning and microscopic interpretation of experiments. Overall, these results highlight how simulations and experiments can complement each other leading to new insights into biomolecular dynamics and function.


[Results of acetabular reconstruction with solid bone graft in primary and revision hip arthroplasty].
To evaluate the results of acetabular reconstruction in total hip replacement and to compare them in view of the use of autologous or allogenic solid bone grafts. The author evaluates a group of 48 total hip replacements carried out in the years 1990-1999, in which solid bone graft was used for reconstruction of an insufficient acetabulum. In 25 hips, the patient's own femoral head was used in primary surgery and, in 23 hips, a frozen femoral head from a tissue bank was used in revision arthroplasty. Most of the acetabular components used in this study were cemented. The patients were evaluated at 5- to 15-year follow-up. Each group was assessed separately and the results were divided into three categories according to X-ray and intra-operative findings, as follows: I. No radiographic evidence of cup loosening and incorporated graft II. Loosened cup and incorporated graft suitable for use in reimplantation III. Loosened cup and graft non-union or resorption In the autograft group, 80 % of the patients fell in category I and 12 % in category II; in the allograft group, 82 % of the patients were in category I and 9 % in category II. Complete failure, i.e., graft resorption and acetabular loosening, was recorded in 8 % and 9 % of group I and group II patients, respectively. Biopsy samples obtained during revision hip surgery showed good incorporation of both autografts and allografts. Other authors' views on the function of solid bone grafts after implantation and on methods of treating a defective acetabulum are discussed. The author puts the good outcomes shown in this study down to a careful implantation technique and thorough preoperative planning. By using a solid graft, the surgeon will achieve good primary fixation of a standard hip socket in a defective acetabulum and will make conditions for repair of defective bone tissue. The results imply that incorporation of both graft and acetabular component can be expected regardless of whether the graft is fresh or frozen. Although this method has its drawbacks and contraindications, it should be considered one of the options when planning total hip arthroplasty for an insufficient acetabulum.


All or none hypothesis: a global-default mode that characterizes the brain and mind.
It is proposed that the mind and brain often work at a gross level and only with fine tuning or inhibition act in a more differentiated manner, even when one might think the domains being issued the global command should be distinct. This applies to disparate findings in cognitive science and neuroscience in both children and adults. Thus, it is easier to switch everything, or nothing, than to switch one thing (the rule one is following or which button to press) but not the other. It is easier to issue the same command to both hands than to move only one hand. If one needs to respond to the opposite (or antonym) of a stimulus, one is faster if the correct response is to the side opposite the stimulus. People tend to think of the nervous system as sending out very precise commands only to the relevant recipient, but it appears that often the command goes out more globally and then parts of the system need to be inhibited from acting on the command.


Force and vibration generated in apical direction by three endodontic files of different kinematics during simulated canal preparation: An in vitro analytical study.
During root canal shaping, pain could result from the high level of force or vibration generated. This could be related to file kinematics or geometry. In the present study, a comparison is made between forces and vibrations generated by endodontic files having three different kinematics. Square pillar resin blocks were used as simulated root canals to study forces and vibrations generated by the file having reciprocating motion (WaveOne Gold), transline motion (Self-Adjusting File), and rotary motion (2Shape). The forces and vibrations were measured using the dynamometer and accelerometer, respectively. Recorded time domain signals were processed in MATLAB to calculate the root mean square value. A one-way analysis of variance and Tukey's test for post hoc comparison at 95% confidence interval were applied over the root mean square data of different files. From a statistical analysis of the file systems, the null hypotheses could not be accepted (P < 0.05) as 95% of the confidence interval. Differences between the means were statistically significant. The root mean square values of force and vibration for WaveOne Gold significantly exceeded those of Self-Adjusting File, 2Shape1, and 2Shape2 while the root mean square values of vibration for 2Shape1 and 2Shape2 were significantly less than the Self-Adjusting File; however, the root mean square value of force for the 2Shape2 was significantly more than for the Self-Adjusting File. Under the present experimental conditions, significant differences in the root mean square values of force and vibration of the three endodontic files of different kinematics have been observed. The WaveOne Gold file system generated higher apical force and vibration than the transline and rotary file system.


[Establishment of a primary culture of cells from embryonic tissue of Loliyo vulgaris L.(Cephalopod)].
The author studies the behaviour in vitro of organ fragments of Loligo vulgaris embryos at hatching stage. These fragments are maintained in three different media. The time of survival of cells is 20 to 25 days in the medium I(sea water), 40 days in the medium II(perivitellin fluid) and 70 to 80 days in the medium II(perivetellin fluid + horse serum). It is this medium III that the original explant is transformed into a typical primary cell culture.


Introduction of pAM beta 1 into Listeria monocytogenes by conjugation and homology between native L. monocytogenes plasmids.
The broad host range antibiotic resistance plasmid pAM beta 1 was transferred from Streptococcus faecalis to 9 of 15 Listeria monocytogenes strains by conjugation. L. monocytogenes transconjugates could transfer the plasmid either among L. monocytogenes strains or back to S. faecalis. Transfer between the various strains occurred without any detectable plasmid DNA rearrangements. The pAM beta 1 replicon was stable in L. monocytogenes--it was retained without antibiotic selection when the bacteria were grown in culture media or passed in mice--and the presence of pAM beta 1 had no major effect on L. monocytogenes virulence. These data suggest that pAM beta 1 or its derivatives might serve as useful L. monocytogenes cloning vehicles. The data presented also demonstrate that pAM beta 1 is compatible with two different native L. monocytogenes plasmids and that Listeria species harbor native plasmids in addition to the 38.5-megadalton plasmid pRYC16 previously reported by Pérez-Díaz et al. (J. C. Pérez-Díaz, M. F. Vicente, and F. Banquero, Plasmid 8:112-118, 1982). Of 29 L. monocytogenes strains screened, 7 contained plasmid DNA. Four strains had similar if not identical plasmids that were 34 megadaltons in size, whereas three other strains contained either a 53-, 44-, or 32-megadalton plasmid; none of these plasmids has the same restriction patterns as pRYC16. DNA homology experiments indicate that the various plasmids are related and suggest that there may be a common set of sequences present in all of the plasmids examined.


Genome-wide analysis reveals unique regulation of transcription of Th2-specific genes by GATA3.
Differentiation of naive CD4 T cells into Th2 cells is accompanied by chromatin remodeling and increased expression of a set of Th2-specific genes, including those encoding Th2 cytokines. IL-4-mediated STAT6 activation induces high levels of transcription of GATA3, a master regulator of Th2 cell differentiation, and enforced expression of GATA3 induces Th2 cytokine expression. However, it remains unclear whether the expression of other Th2-specific genes is induced directly by GATA3. A genome-wide unbiased chromatin immunoprecipitation assay coupled with massive parallel sequencing analysis revealed that GATA3 bound to 1279 genes selectively in Th2 cells, and 101 genes in both Th1 and Th2 cells. Simultaneously, we identified 26 highly Th2-specific STAT6-dependent inducible genes by DNA microarray analysis-based three-step selection processes, and among them 17 genes showed GATA3 binding. We assessed dependency on GATA3 for the transcription of these 26 Th2-specific genes, and 10 genes showed increased transcription in a GATA3-dependent manner, whereas 16 genes showed no significant responses. The transcription of the 16 GATA3-nonresponding genes was clearly increased by the introduction of an active form of STAT6, STAT6VT. Therefore, although GATA3 has been recognized as a master regulator of Th2 cell differentiation, many Th2-specific genes are not regulated by GATA3 itself, but in collaboration with STAT6.


Trends in male contraception.
Methods that are available for male contraception, namely coitus interruptus, condoms, and vasectomy, have been used since the 19th century. With the exceptions of a few improvements of these methods, no major progress has been made with respect to introducing new male contraceptives since then. It is extremely urgent to develop new, safe, effective, and reversible male contraceptive methods. Among all male contraceptive methods that are being investigated, the hormonal approach is the closest to clinical application. Hormonal contraception provides pregnancy protection by means of spermatogenic suppression. Androgen-progestin regimens currently represent the best available hormonal combination for induction of a profound suppression of spermatogenesis. Further development of new steroids is mandatory for increasing the choices of available contraceptive formulations and to optimize long-term safety of these regimens.


Pollination in the Chilean Mediterranean-type ecosystem: a review of current advances and pending tasks.
We conducted a systematic review of the scientific literature published on plant-pollinator interactions, from both the plant and pollinator perspective, in the Chilean Mediterranean-type ecosystem (MTE hereafter). Our search identified 69 published papers on 235 native plant species from 62 families. Less than 7.9% of the flowering species inhabiting the Chilean Mediterranean have been studied, and most studies were restricted to only one locality and one reproductive season. The geographic location of the studies differed from a random pattern, showing two well-defined areas where most studies were conducted. Likewise, most studies in the Andes Range were performed above 2000 m a.s.l. The number of species of flower visitor per plant species was low (4.25 ± 0.22), which probably results from the historical and biogeographic isolation of Chile. This literature survey shows that studies relating floral traits with pollinator attraction and plant reproduction are the most frequent topics of research, reaching 37.6% of studies, followed by studies that examine pollination in relation to human impact (16.1%), micro- and macroevolution (14.0%), relationships between pollination and other ecological interactions (10.8%), community and network assessments (11.8%), and effects of abiotic variables on pollination interactions (9.7%). Our review highlights a lack of research on the effects of pollination for anthropogenic land use especially as agricultural practice is one of the most salient features of the Chilean MTE. Future directions to increase our understanding of the role of plant-pollinator relationships for biodiversity maintenance should include: to extend the taxonomic and geographic scope of research, to increase the number of spatial and temporal replicates, to increase the number of studies on pollination networks as they provide estimates of community complexity and putative stability, to develop studies that estimate the importance of pollination for plant demographic parameters and conservation, and to conduct studies that estimate the ecological service provided by Chilean native pollinators for crop yield and sustainable agriculture.


The social motivation theory of autism.
The idea that social motivation deficits play a central role in Autism Spectrum Disorders (ASD) has recently gained increased interest. This constitutes a shift in autism research, which has traditionally focused more intensely on cognitive impairments, such as theory-of-mind deficits or executive dysfunction, and has granted comparatively less attention to motivational factors. This review delineates the concept of social motivation and capitalizes on recent findings in several research areas to provide an integrated account of social motivation at the behavioral, biological and evolutionary levels. We conclude that ASD can be construed as an extreme case of diminished social motivation and, as such, provides a powerful model to understand humans' intrinsic drive to seek acceptance and avoid rejection.


Rethinking Intrusiveness: Exploring the Sequential Organization in Interactions Between Infants and Mothers.
To date, studies investigating maternal postpartum depression (PPD) have mainly focused on identifying failures in interactions of postpartum depressed mothers and their infants, often attributed to single dysfunctional maternal behaviors. Intrusiveness has been identified as a dysfunctional behavior characterizing mothers suffering from PPD. However, this research does not consider the co-constructed and sequential nature of social interactions, in which single behaviors cannot be conceived as isolated or disconnected units. The aim of the work presented in this paper was to explore the interactional dynamics underlying maternal behaviors previously identified as intrusive by mainstream literature on postpartum depression. Through a conversation analytical approach, we analyzed filmed interactions between mothers with and without postpartum depression and their 3-months-old infants. The analyses of 4 selected episodes illustrate similar dyadic activities, yet presenting different levels of mutuality and affective attunement. Results showed two normative features of social interactions that contributed to the different quality in the mutual adjustment of the partners: interactional rhythm and preliminaries. Interactional rhythm refers to the structuring of infants' spontaneous activity into a turn sequence, whereas preliminaries consist of verbal or nonverbal moves that anticipate following action. As evident from our analytical observations, what seems to be hindering the mutual coordination (previously labeled as "intrusive") is not based on specific individual behaviors but on the absence or violation of such interactional norms. Adopting an interactive and dynamical framework, we shifted the focus from maternal behaviors considered as dysfunctional to observing the unfolding of interactional aspects contributing to better or poorer sequential structuring. We argue that these aspects shape the possibilities for the infant's participation. Finally, we discuss the theoretical and methodological implications of adopting a conversation analytical approach for a better understanding of the relational dynamics related to clinical and non-clinical interactions.


Accuracy of 3 Tesla pelvic phased-array multiparametric MRI in diagnosing prostate cancer at repeat biopsy.
Multiparametric pelvic magnetic resonance imaging (mpMRI) accuracy in prostate cancer (PCa) diagnosis was evaluated. From June 2011 to December 2013, 168 patients (median 65 years) with negative digital rectal examination underwent repeat transperineal saturation biopsy (SPBx; median 28 cores) for persistently high or increasing PSA values, PSA >10 ng/ml or PSA values between 4.1-10 o r 2.6-4 ng/ml with free/total PSA < 25% and < 20%, respectively. All patients underwent mpMRI using a 3.0 Tesla scanner equipped with surface 16 channels phased-array coil and lesions suspicious for PCa were submitted to additional targeted biopsies. A T1c PCa was found in 66 (39%) cases; SPBx and mpMRI-suspicious targeted biopsy diagnosed 60 (91%) and 52 (78.8%) cancers missing 6 (all of the anterior zone) and 14 cancers (12 and 2 of the lateral margins and anterior zone), respectively; in detail, mpMRI missed 12 (18.1%) PCa charaterized by microfocal (1 positive core with greatest percentage of cancer and Gleason score equal to 5% and 6, respectively) disease at risk for insignificant cancer. The diameter of the suspicious mpMRI lesion was directly correlated to the diagnosis of PCa with poor Gleason score (p < 0.05); detection rate of cancer for each suspicious mpMRI core was 35.3%. Diagnostic accuracy, sensitivity, specificity, positive and negative predictive value of mpMRI in diagnosing PCa was 75.7%, 82.5%, 71.8%, 78.9%, 87.9%, respectively. Multiparametric pMRI improved SPBx accuracy in diagnosing significant anterior PCa; the diameter of mpMRI suspicious lesion resulted significantly predictive of aggressive cancers.


Muscle synergies involved in shifting the center of pressure while making a first step.
We used the framework of the uncontrolled manifold (UCM) hypothesis to analyze multi-muscle synergies involved in making a step by a standing person. We hypothesized that leg and trunk muscles are organized into stable groups (muscle modes, M-modes) related to shifts of the center of pressure (COP) in the anterior-posterior and medio-lateral directions. Another hypothesis was that the magnitudes of the modes co-vary across repetitive trials to stabilize a certain magnitude of the COP shift in both directions. M-modes were defined using principal component analysis applied to indices of changes in the electromyographic (EMG) activity prior to releasing variable loads that were held by the subject using a pulley system. For the task of releasing the load behind the body three M-modes associated with a backward COP shift were defined. Four M-modes were defined for the task of releasing the load at the body side associated with a lateral COP shift. Multiple regression analysis was used to relate changes in the M-mode magnitudes to COP shifts. EMG changes prior to making a step were quantified over five 100 ms time windows before the lift-off of the stepping leg. Two components of the variance in the M-mode space computed across repetitions of a stepping task were quantified-a component that did not affect the average COP shift in a particular direction (variance within the UCM, V (UCM)), and a component that affected the COP shift (variance orthogonal to the UCM, V (ORT)). V (UCM) was significantly higher than V (ORT) for both directions of the COP shifts. This relation was observed for the M-modes in the stepping leg as well as in the support leg. The stepping leg showed a different time evolution of the ratio V (UCM)/V (ORT) such that the difference between the two variance components disappeared closer to the time of the lift-off. The findings corroborate both main hypotheses. The study supports a view that control of whole-body actions involves grouping the muscles, using fewer elemental variables to scale the muscle activity, and forming synergies in the space of the elemental variables that stabilize time profiles of important performance variables.


Congenital hyperinsulinism in Brazilian neonates: a study of histology, KATP channel genes, and proliferation of β cells.
Congenital hyperinsulinism (CHI) is a rare pancreatic β-cell disease of neonates, characterized by inappropriate insulin secretion with severe persistent hypoglycemia, with regard to which many questions remain to be answered, despite the important acquisition of its molecular mechanisms in the last decade. The aim of this study was to examine pancreatic histology, β-cell proliferation (immunohistochemistry with double staining for Ki-67/insulin), and β-cell adenosine triphosphate-sensitive potassium channels genes from 11 Brazilian patients with severe medically unresponsive CHI who underwent pancreatectomy. Pancreatic histology and β-cell proliferation in CHI patients were compared to pancreatic samples from 19 age-matched controls. Ten cases were classified as diffuse form (D-CHI) and 1 as focal form (F-CHI). β-cell nucleomegaly and abundant cytoplasm were absent in controls and were observed only in D-CHI patients. The Ki-67 labeling index (Ki-67-LI) was used to differentiate the adenomatous areas of the F-CHI case (10.15%) from the "loose cluster of islets" found in 2 D-CHI samples (2.29% and 2.43%) and 1 control (1.54%) sample. The Ki-67-LI was higher in the F-CHI adenomatous areas, but D-CHI patients also had significantly greater Ki-67-LI (mean value = 2.41%) than age-matched controls (mean value = 1.87%) (P = 0.009). In this 1st genetic study of CHI patients in Brazil, no mutations or new polymorphisms were found in the 33-37 exons of the ABCC8 gene (SUR1) or in the entire exon of the KCNJ11 gene (Kir 6.2) in 4 of 4 patients evaluated. On the other hand, enhanced β-cell proliferation seems to be a constant feature in CHI patients, both in diffuse and focal forms.


Microbial translocation: translating simian immunodeficiency virus to HIV.
The article describes recent advances in understanding the causes and consequences of microbial translocation in HIV and simian immunodeficiency virus infections. Persistent microbial translocation contributes to aberrant immune activation in immunodeficiency lentiviral infections and thereby, pathogenesis and mortality. Efforts to delineate the circumstances surrounding translocation have benefited from use of simian immunodeficiency virus-infected nonhuman primates and highlight the overwhelming immunologic diversion caused by translocating microbes. The use of therapeutics aimed at reducing microbial translocation show promise and will benefit from continued research into the mechanisms that promote systemic microbial dissemination in treated and untreated infections. Insights into the source and identity of translocating microbes in lentiviral infections continue to enhance the development of adjunct therapeutics.


The effect of metformin on TSH levels in euthyroid and hypothyroid newly diagnosed diabetes mellitus type 2 patients.
Metformin is the first-line oral hypoglycemic agent in the treatment of type 2 diabetes mellitus with a number of positive effects. The aim of the study was to determine the effect of metformin on TSH levels in euthyroid and hypothyroid newly diagnosed diabetes mellitus type 2 patients. The study included 255 newly diagnosed diabetes mellitus type 2 drug naive patients, 170 euthyroid patinets, group A, 85 hypothiroid patients, group B, and 80 euthyroid DM type 2 patients on metformin therapy for more than 5 years, group C. Patients in groups A and B began metformin treatment with a dose of 2000 mg/day. We assessed baseline TSH, FT3, FT4 levels and TPOab, in groups A , B and C, and 6 months after initiation of metformin therapy in groups A and B. There were no differences in FT3 and FT4 levels after 6 months of metformin treatment in all groups. TSH level in Group A showed some reduction after 6 months of metformin therapy, not statistically significant. The only statistically significant change in Group A is the change of TSH level after 6 months in TPOAb positive patients. There was statiscically significant decrease in TSH level after 6 months in group B. There were no significant differences of basal TSH, FT3 and FT4 levels in groups A and B compared to group C. The results show that metformin has TSH lowering effect in patients with type 2 DM and hypothyreoidism, but also in euthyroid TPOab positive, levothyroxine naive patients. We have shown that the TSH lowering effect of metformin is not dependent on long term metformin therapy (Tab. 2, Ref. 18).


Classification of Sport-Related Head Trauma: A Spectrum of Mild to Severe Injury.
OBJECTIVE: To identify the types of injuries the human brain incurs as a result of traumatic forces applied to the cranium. In athletic events and endeavors, the full spectrum of intracranial hemorrhages in various compartments, raised intracranial pressure, and diffuse nonhemorrhagic damage may be seen. In this review, we describe these serious injuries and the more common mild traumatic brain injury in their clinical presentations and relate concussion classification to the overall picture of traumatic brain injury. METHODS: Our cumulative experience with athletic injuries, both at the catastrophic and mild traumatic brain injury levels, has led us to a management paradigm that serves to guide us in the classification and treatment of these athletes. DISCUSSION: The occurrence of intracranial injuries in sports has now been well documented. Intracranial hematomas (epidural, subdural, and parenchymal) and cerebral contusions can result from head injury. Many patients sustain a diffuse brain injury, resulting in elevated intracranial pressures, without a blood clot or mass lesion. The classification of concussion and the use of concussion guidelines are not uniform. However, the major emphases are agreed upon: the close and careful scrutiny of the athlete, an expeditious but reliable neurologic examination, and proper on-field management. Return-to-play decisions are based on many factors that affect normal functioning, both on and off the playing field. CONCLUSIONS: Sufficient knowledge now exists to allow us to carefully evaluate the injured athlete, to place him or her in the management scheme to minimize the potential for permanent cerebral dysfunction, and to know when the athlete can safely return to contact sport participation.


Androgen resistance in squirrel monkeys (Saimiri spp.).
The goal of this study was to understand the basis for high androgen levels in squirrel monkeys (Saimiri spp.). Mass spectrometry was used to analyze serum testosterone, androstenedione, and dihydrotestosterone of male squirrel monkeys during the nonbreeding (n = 7) and breeding (n = 10) seasons. All hormone levels were elevated compared with those of humans, even during the nonbreeding season; the highest levels occurred during the breeding season. The ratio of testosterone to dihydrotestosterone in squirrel monkeys is high during the breeding season compared to man. Squirrel monkeys may have high testosterone to compensate for inefficient metabolism to dihydrotestosterone. We also investigated whether squirrel monkeys have high androgens to compensate for low-activity androgen receptors (AR). The response to dihydrotestosterone in squirrel monkey cells transfected with AR and AR-responsive reporter plasmids was 4-fold, compared with 28-fold in human cells. This result was not due to overexpression of cellular FKBP51, which causes glucocorticoid and progestin resistance in squirrel monkeys, because overexpression of FKBP51 had no effect on dihydrotestosterone-stimulated reporter activity in a human fibroblast cell line. To test whether the inherently low levels of FKBP52 in squirrel monkeys contribute to androgen insensitivity, squirrel monkey cells were transfected with an AR expression plasmid, an AR-responsive reporter plasmid, and a plasmid expressing FKBP52. Expression of FKBP52 decreased the EC50 or increased the maximal response to dihydrotestosterone. Therefore, the high androgen levels in squirrel monkeys likely compensate for their relatively low 5 alpha-reductase activity during the breeding season and AR insensitivity resulting from low cellular levels of FKBP52.


Interactions of fibrinolytic system proteins with lysine-containing surfaces.
Studies on the interactions of tissue plasminogen activator (tPA) and plasminogen with polyurethane surfaces containing epsilon-lysine moieties (epsilon-amino group free) are reported. These surfaces are considered to have the potential to dissolve nascent clots that may be formed on them. For adsorption from both single protein solutions and plasma, the surfaces were found to have a high capacity for tPA as well as plasminogen. A significant fraction of preadsorbed tPA was displaced from the epsilon-lysine surfaces upon contact with plasma. These surfaces, when preadsorbed with tPA and then incubated with plasma, were able to dissolve incipient clots formed around them. However, the clot-dissolving capacity diminished as the time of plasma incubation increased, presumably due to loss of tPA. It was also shown that in plasma, preadsorbed tPA is displaced from these surfaces largely by plasminogen, which thus appears to have a greater binding affinity than tPA for the epsilon-lysine moieties. Finally, it was found that in plasma, the epsilon-lysine surfaces interact with plasminogen in a dynamic manner, and that about 70% of the bound plasminogen is exchanging continuously with plasminogen in the plasma.


Diabetes Mellitus and Heart Failure.
Epidemiologic and clinical data from the last 2 decades have shown that the prevalence of heart failure in diabetes is very high, and the prognosis for patients with heart failure is worse in those with diabetes than in those without diabetes. Experimental data suggest that various mechanisms contribute to the impairment in systolic and diastolic function in patients with diabetes, and there is an increased recognition that these patients develop heart failure independent of the presence of coronary artery disease or its associated risk factors. In addition, current clinical data demonstrated that treatment with the sodium glucose cotransporter 2 inhibitor empagliflozin reduced hospitalization for heart failure in patients with type 2 diabetes mellitus and high cardiovascular risk. This review article summarizes recent data on the prevalence, prognosis, pathophysiology, and therapeutic strategies to treat patients with diabetes and heart failure.


Reproducibility of heart rate recovery in patients with intermittent claudication.
Postexercise heart rate recovery (HRR) is a non-invasive tool for cardiac autonomic function assessment. Reproducibility of HRR has been established in healthy subjects; however, no study has evaluated this reproducibility in clinical populations who may present autonomic dysfunction. Patients with peripheral artery disease and intermittent claudication (IC) often present altered cardiac autonomic function and HRR could be an interesting tool for evaluating autonomic responses to interventions in this population. Therefore, the reproducibility of HRR should be determined in this specific population. To determine the reproducibility of HRR indices in patients with IC. Nineteen men with IC underwent two repeated maximal treadmill tests. Raw HR and relative HRR (difference to exercise peak) indices measured at 30, 60, 120, 180, 240 and 300s of recovery were evaluated. The presence of systematic bias was assessed by comparing test and retest mean values via paired t-test. Reliability was assessed by intraclass correlation coefficient (ICC), and agreement by typical error (TE), coefficient of variation (CV) and minimal detectable difference (MDD). There were no significant differences between the test and retest values of all raw HR and relative HRR indices (P ≥ 0·05), except for HR120s (P = 0·032). All indices exhibited excellent reliability (ICC ≥ 0·78). Raw HR and relative HRR indices showed TEs ≤ 6·4 bpm and MDDs ≤ 17·8 bpm. In addition, all indices showed CVs ≤ 13·2%, except HRR30s (CV = 45·6%). The current results demonstrated that most HRR indices were highly reproducible with no systematic error, excellent reliability and good agreement in patients with IC following maximal graded exercise.


A health equity framework to support the next generation of cancer population simulation models.
Over the past 2 decades, population simulation modeling has evolved as an effective public health tool for surveillance of cancer trends and estimation of the impact of screening and treatment strategies on incidence and mortality, including documentation of persistent cancer inequities. The goal of this research was to provide a framework to support the next generation of cancer population simulation models to identify leverage points in the cancer control continuum to accelerate achievement of equity in cancer care for minoritized populations. In our framework, systemic racism is conceptualized as the root cause of inequity and an upstream influence acting on subsequent downstream events, which ultimately exert physiological effects on cancer incidence and mortality and competing comorbidities. To date, most simulation models investigating racial inequity have used individual-level race variables. Individual-level race is a proxy for exposure to systemic racism, not a biological construct. However, single-level race variables are suboptimal proxies for the multilevel systems, policies, and practices that perpetuate inequity. We recommend that future models designed to capture relationships between systemic racism and cancer outcomes replace or extend single-level race variables with multilevel measures that capture structural, interpersonal, and internalized racism. Models should investigate actionable levers, such as changes in health care, education, and economic structures and policies to increase equity and reductions in health-care-based interpersonal racism. This integrated approach could support novel research approaches, make explicit the effects of different structures and policies, highlight data gaps in interactions between model components mirroring how factors act in the real world, inform how we collect data to model cancer equity, and generate results that could inform policy.


Protective effects of the fermented milk Kefir on X-ray irradiation-induced intestinal damage in B6C3F1 mice.
Gastrointestinal damage associated with radiation therapy is currently an inevitable outcome. The protective effect of Kefir was assessed for its usefulness against radiation-induced gastrointestinal damage. A Kefir supernatant was diluted by 2- or 10-fold and administered for 1 week prior to 8 Gray (Gy) X-ray irradiation at a dose rate of 2 Gy/min, with an additional 15 d of administration post-irradiation. The survival rate of control mice with normal drinking water dropped to 70% on days 4 through 9 post-irradiation. On the other hand, 100% of mice in the 10- and 2-fold-diluted Kefir groups survived up to day 9 post-irradiation (p<0.05 and p<0.01, respectively). Examinations for crypt regeneration against 8, 10 and 12 Gy irradiation at a dose rate of 4 Gy/min revealed that the crypt number was significantly increased in the mice administered both diluted Kefir solutions (p<0.01 for each). Histological and immunohistochemical examinations revealed that the diluted Kefir solutions protected the crypts from radiation, and promoted crypt regeneration. In addition, lyophilized Kefir powder was found to significantly recover the testis weights (p<0.05), but had no effects on the body and spleen weights, after 8 Gy irradiation. These findings suggest that Kefir could be a promising candidate as a radiation-protective agent.


Diffusion MRI simulation in thin-layer and thin-tube media using a discretization on manifolds.
The Bloch-Torrey partial differential equation can be used to describe the evolution of the transverse magnetization of the imaged sample under the influence of diffusion-encoding magnetic field gradients inside the MRI scanner. The integral of the magnetization inside a voxel gives the simulated diffusion MRI signal. This paper proposes a finite element discretization on manifolds in order to efficiently simulate the diffusion MRI signal in domains that have a thin layer or a thin tube geometrical structure. The variable thickness of the three-dimensional domains is included in the weak formulation established on the manifolds. We conducted a numerical study of the proposed approach by simulating the diffusion MRI signals from the extracellular space (a thin layer medium) and from neurons (a thin tube medium), comparing the results with the reference signals obtained using a standard three-dimensional finite element discretization. We show good agreements between the simulated signals using our proposed method and the reference signals for a wide range of diffusion MRI parameters. The approximation becomes better as the diffusion time increases. The method helps to significantly reduce the required simulation time, computational memory, and difficulties associated with mesh generation, thus opening the possibilities to simulating complicated structures at low cost for a better understanding of diffusion MRI in the brain.


Neonatal Colonic Inflammation Epigenetically Aggravates Epithelial Inflammatory Responses to Injury in Adult Life.
Early life adversity is considered a risk factor for the development of gastrointestinal diseases, including inflammatory bowel disease. We hypothesized that early life colonic inflammation causes susceptibility to aggravated overexpression of interleukin (IL)1β. We developed a 2-hit rat model in which neonatal inflammation (NI) and adult inflammation (AI) were induced by trinitrobenzene sulfonic acid. Aggravated immune responses were observed in NI + AI rats, including a sustained up-regulation of IL1β and other cytokines. In parallel with exacerbated loss of inhibitor of kappa B alpha expression, NI + AI rats showed hyperacetylation of histone H4K12 and increased V-Rel Avian Reticuloendotheliosis Viral Oncogene Homolog A binding on the IL1B promoter, accompanied by high levels of norepinephrine/epinephrine. Propranolol, a β-blocker, markedly ameliorated the inflammatory response and IL1β overexpression by mitigating against epigenetic modifications. Adrenalectomy abrogated NI-induced disease susceptibility whereas yohimbine sensitized the epithelium for exacerbated immune response. The macrophages of NI rats produced more IL1β than controls after exposure to lipopolysaccharide (LPS), suggesting hypersensitization; incubation with LPS plus Foradil (Sigma, St. Louis, MO), a β2-agonist, induced a greater IL1β expression than LPS alone. Epinephrine and Foradil also exacerbated LPS-induced IL1β activation in human THP-1-derived macrophages, by increasing acetylated H4K12, and these increases were abrogated by propranolol. NI sensitizes the colon epithelium for exacerbated IL1β activation by increasing stress hormones that induce histone hyperacetylation, allowing greater access of nuclear factor-κB to the IL1B promoter and rendering the host susceptible to aggravated immune responses. Our findings suggest that β blockers have a therapeutic potential for inflammatory bowel disease susceptibility and establish a novel paradigm whereby NI induces epigenetic susceptibility to inflammatory bowel disease.


Molecule imprinting chiral stationary phase.
Molecule imprinting polymers (MIPs) with high chiral selectivity for N(alpha)-protected amino acids were synthesized in polar solvent using acrylamide (AM) or combined functional monomer methacrylic acid (MAA) + vinyl pyridine (VP). Factors that influence the chiral selectivity of MIPs and mechanisms of the chiral recognition process were investigated. It was found that the rigid structure and the polar functional group of the print molecule were very important for the chiral selectivity of MIPs. For MIPs made using a combined functional monomer, ionic and hydrophobic interaction may be responsible for the chiral recognition process in aqueous media. The number of binding sites on MIPs and dissociation constants for the complex of enantiomers and MIPs were determined by frontal chromatography analysis.


A prospective randomized trial of single- or dual-chamber implantable cardioverter-defibrillators to minimize inappropriate shock risk in primary sudden cardiac death prevention.
Dual-chamber implantable cardioverter-defibrillators (ICDs) may improve specificity and reduce the risk of inappropriate shocks, and enhance atrial arrhythmia (AT/AF) detection to permit stroke prevention compared with single-chamber ICDs, but at additional expense and risk. Patients (n = 100) receiving primary prevention ICDs at two USA and two Israeli centres were randomized to dual-chamber or single-chamber devices between December 2008 and December 2010 and were followed for 1 year. Programming in both groups included: delayed detection to avoid therapy for non-sustained episodes; high detection cut-off rates to avoid treating slower, better tolerated arrhythmias; minimized right ventricular pacing; and routine use of supraventricular-ventricular tahcycardia discriminators and antitachycardia pacing. The primary outcome was the proportion of patients with inappropriate shocks. One patient in each group (2%) received inappropriate shocks (P = 1.00). Death occurred in two patients in the single-chamber arm, and in none of the patients in the dual-chamber arm (P = 0.15). New AT/AF was detected in 12 patients (24%) in the dual-chamber group, vs. no patients in the single-chamber group (P < 0.001). Among US participants, the mean cost of dual- vs. single-chamber ICD implantation was $16 579 vs. $14 249, respectively (P < 0.001); there was no difference in the quality of life (EQ-5D index difference 0.013, P = 0.769; EQ VAS difference 3.3, P = 0.49). When optimal programming is utilized, inappropriate shocks are rare in primary prevention patients with both single- and dual-chamber ICDs. The routine use of dual-chamber ICDs increases the expense without reducing inappropriate shocks or improving the quality of life at 1 year. clinicaltrials.gov Identifier: NCT00787800.


Prolonged and recurrent occupational dermatitis. Some whys and wherefores.
Most cases of occupational dermatitis are successfully diagnosed and treated effectively; with proper preventive measures, the patient can resume work. This is not always the case, however, because there are certain employees whose dermatitis defies the best diagnostic and therapeutic measures. Such cases engender disagreement between the patient and physician, the employer, the insurance carrier, and the compensation commission. Several of the causes for failure to effect a cure in cases of occupational dermatitis are explained in this article.


[Structure of the DNA molecule in the course of reparation process after ultraviolet irradiation of E. coli cells].
Kinetics was studied of DNA degradation, processing and synthesis in the course of post-radiation incubation (irradiation dose 0-2000 erg/mm2). It has been shown that contrary to the endonuclease splitting of DNA strands, DNA degradation requires energy. DNA degradation is not enhanced upon an increase of the irradiation up to 400 erg/mm2 and more and at the incubation duration exceeding 120 min. The degradation involves not more than 40% of the total DNA quantity. It takes place in the absence of resynthesis registered by the incorporation of [3h] thymidine and 32P and dephinylamine reaction. The number of single-strand breaks in DNA (per one E. coli chromosome strans) reaches a maximum value after 120-180 min. of incubation. This maximal value is linearily increased upon an increase of the irradiation dose from 0 to 1200 erg/mm2 and does not change when the dose increases further. The number of single-strand breaks does not exceed 35-50, this value being dependent on the presence of glucose. On the basis of the data obtained a suggestion is put forward that in the course of the reparative degradation of DNA, prolonged (10(4)-10(5) nucleotides) single-stranded regions are formed. Possible role of these 'nicks" in the appearance of mutations is discussed.


[Acute ethylene glycol poisoning].
During last 7 years, 13 cases of acute ethylene glycol poisoning have been observed with 4 fatal outbreaks. All cases--except one--were accidental. Six patients had drunk a mixture of antifreeze in water as they were lost in a desert. A patient who had taken 970 ml of ethylene glycol survived. In one case, death was due to irreversible brain damage; two other fatalities occurred from cardiorespiratory distress. CNS involvement was noticed in 8 cases and acute renal failure-constantly controlled-occurred in 9 patients. Post mortem examination has shown bi-refringent calcium oxalate crystals in both kidney and brain. Specific treatment with ethanol has been performed in 3 cases with ingestion of large amounts of toxic who were seen before definitive renal lesions have occurred. Emphasis is placed in symptomatic treatment including gastric lavage, extrarenal epuration and conservative management.


Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties.
The cancer stem cell theory hypothesizes that cancer stem cells (CSCs), which possess self-renewal and other stem cell properties, are regarded as the cause of tumor formation, recurrence and metastasis. The isolation and identification of CSCs could help to develop novel therapeutic strategies specifically targeting CSCs. In this study, we enriched gastric cancer stem cells through spheroid body formation by cultivating the human gastric cancer cell line MKN-45 in defined serum-free medium. The stemness characteristics of spheroid body-forming cells, including self-renewal, proliferation, chemoresistance, tumorigenicity of the MKN-45 spheroid body-forming cells were evaluated, and the expression levels of stemness genes and related proteins in the MKN-45 spheroid body-forming cells were assessed. Furthermore, immunofluorescence staining for the stem cell markers on spheroid body-forming cells was examined to evaluate the association between stemness factors (Oct4, Sox2, Nanog) and the proposed CSC marker CD44. Our data demonstrated that non-adherent spheroid body-forming cells from the gastric cancer cell line MKN-45 cultured in stem cell-conditioned medium possessed gastric CSC properties, such as persistent self-renewal, extensive proliferation, drug resistance, high tumorigenic capacity and overexpression of CSC-related genes and proteins (Oct4, Sox2, Nanog and CD44), compared with the parental cells. More importantly, CD44-positive cells co-expressing the pluripotency genes Oct4, Sox2 and Nanog may represent gastric CSCs. Further experiments using more refined selection criteria such as a combination of two or multiple markers would be useful to specifically identify and purify CSCs.


[Effect of quinine on the calcium currents of mollusk neurons].
The effects of quinine on the peak amplitude and relaxation of calcium current (ICa) were examined on isolated unidentified snail neurons. Quinine (1 X 10(-5)-5 X 10(-4) mol/l) reversibly slowed the relaxation of ICa in a dose-dependent manner (early effect) and suppressed the peak amplitude of ICa in a dose-dependent and time-dependent manner (delayed effect). Quinine induced a shift of the current-voltage relationship for peak ICa by 5-10 mV in the hyperpolarizing direction. A half inhibition of ICa peak amplitude was induced by 6 X 10(-5) mol/l quinine. The results show that quinine is an effective inhibitor of calcium channels in snail neurons.


Medullary substrate and differential cardiovascular responses during stimulation of specific acupoints.
Electroacupuncture (EA) at P5-P6 acupoints overlying the median nerve reduces premotor sympathetic cardiovascular neuronal activity in the rostral ventral lateral medulla (rVLM) and visceral reflex pressor responses. In previous studies, we have noted different durations of influence of EA comparing P5-P6 and S36-S37 acupoints, suggesting that point specificity may exist. The purpose of this study was to evaluate the influence of stimulating P5-P6 (overlying the median nerve), LI4-L7 (overlying branches of the median nerve and the superficial radial nerve), LI6-LI7 (overlying the superficial radial nerve), LI10-LI11 (overlying the deep radial nerves), S36-S37 (overlying the deep peroneal nerves), or K1-B67 (overlying terminal branches of the tibial nerves) specific acupoints, overlying deep and superficial somatic nerves, on the excitatory cardiovascular reflex and rVLM responses evoked by stimulation of chemosensitive receptors in the cat's gallbladder with bradykinin (BK) or direct splanchnic nerve (SN) stimulation. We observed point-specific differences in magnitude and duration of EA inhibition between P5-P6 or LI10-LI11 and LI4-L7 or S36-S37 in responses to 30-min stimulation with low-frequency, low-current EA. EA at LI6-LI7 and K1-B67 acupoints as well as direct stimulation of the superficial radial nerve did not cause any cardiovascular or rVLM neuronal effects. Cardiovascular neurons in the rVLM, a subset of which were classified as premotor sympathetic cells, responded to brief (30 s) stimulation of the SN as well as acupoints P5-P6, LI10-LI11, LI4-L7, S36-S37, LI6-LI7, or K1-B67, or underlying somatic pathways in a fashion similar to the reflex responses. In fact, we observed a significant linear relationship (r(2) = 0.71) between the evoked rVLM response and reflex change in mean arterial blood pressure. In addition, EA stimulation at P5-P6 and LI4-L7 decreased rVLM neuronal activity by 41 and 12%, respectively, for >1 h, demonstrating that prolonged input into the medulla during stimulation of somatic nerves, depending on the degree of convergence, leads to more or less inhibition of activity of these cardiovascular neurons. Thus EA at acupoints overlying deep and superficial somatic nerves leads to point-specific effects on cardiovascular reflex responses. In a similar manner, sympathetic cardiovascular rVLM neurons that respond to both visceral (reflex) and somatic (EA) nerve stimulation manifest graded responses during stimulation of specific acupoints, suggesting that this medullary region plays a role in site-specific inhibition of cardiovascular reflex responses by acupuncture.


Oxidized low-density lipoproteins as a novel risk factor and therapeutic target for ACM.
Arrhythmogenic cardiomyopathy (ACM) is an inherited heart disease involving arrhythmia in young adults accompanied by structural changes at later stages. In this issue of EMBO Molecular Medicine, Sommariva et al (2021) identified a positive correlation between circulating levels of oxidized low-density lipoproteins (oxLDL) and ACM disease penetrance, which contributes to fibro-fatty cardiac remodeling via the oxLDL/CD36/PPARγ axis. These data identify oxidized low-density lipoproteins as a risk factor for ACM and uncover a novel therapeutic intervention option to block disease pathogenesis.


Analysis of non-TIR NBS-LRR resistance gene analogs in Musa acuminata Colla: isolation, RFLP marker development, and physical mapping.
Many commercial banana varieties lack sources of resistance to pests and diseases, as a consequence of sterility and narrow genetic background. Fertile wild relatives, by contrast, possess greater variability and represent potential sources of disease resistance genes (R-genes). The largest known family of plant R-genes encode proteins with nucleotide-binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. Conserved motifs in such genes in diverse plant species offer a means for isolation of candidate genes in banana which may be involved in plant defence. A computational strategy was developed for unbiased conserved motif discovery in NBS and LRR domains in R-genes and homologues in monocotyledonous plant species. Degenerate PCR primers targeting conserved motifs were tested on the wild cultivar Musa acuminata subsp. burmannicoides, var. Calcutta 4, which is resistant to a number of fungal pathogens and nematodes. One hundred and seventy four resistance gene analogs (RGAs) were amplified and assembled into 52 contiguous sequences. Motifs present were typical of the non-TIR NBS-LRR RGA subfamily. A phylogenetic analysis of deduced amino-acid sequences for 33 RGAs with contiguous open reading frames (ORFs), together with RGAs from Arabidopsis thaliana and Oryza sativa, grouped most Musa RGAs within monocotyledon-specific clades. RFLP-RGA markers were developed, with 12 displaying distinct polymorphisms in parentals and F1 progeny of a diploid M. acuminata mapping population. Eighty eight BAC clones were identified in M. acuminata Calcutta 4, M. acuminata Grande Naine, and M. balbisiana Pisang Klutuk Wulung BAC libraries when hybridized to two RGA probes. Multiple copy RGAs were common within BAC clones, potentially representing variation reservoirs for evolution of new R-gene specificities. This is the first large scale analysis of NBS-LRR RGAs in M. acuminata Calcutta 4. Contig sequences were deposited in GenBank and assigned numbers ER935972 - ER936023. RGA sequences and isolated BACs are a valuable resource for R-gene discovery, and in future applications will provide insight into the organization and evolution of NBS-LRR R-genes in the Musa A and B genome. The developed RFLP-RGA markers are applicable for genetic map development and marker assisted selection for defined traits such as pest and disease resistance.


Comparative transcriptome analysis of obligately asexual and cyclically sexual rotifers reveals genes with putative functions in sexual reproduction, dormancy, and asexual egg production.
Sexual reproduction is a widely studied biological process because it is critically important to the genetics, evolution, and ecology of eukaryotes. Despite decades of study on this topic, no comprehensive explanation has been accepted that explains the evolutionary forces underlying its prevalence and persistence in nature. Monogonont rotifers offer a useful system for experimental studies relating to the evolution of sexual reproduction due to their rapid reproductive rate and close relationship to the putatively ancient asexual bdelloid rotifers. However, little is known about the molecular underpinnings of sex in any rotifer species. We generated mRNA-seq libraries for obligate parthenogenetic (OP) and cyclical parthenogenetic (CP) strains of the monogonont rotifer, Brachionus calyciflorus, to identify genes specific to both modes of reproduction. Our differential expression analysis identified receptors with putative roles in signaling pathways responsible for the transition from asexual to sexual reproduction. Differential expression of a specific copy of the duplicated cell cycle regulatory gene CDC20 and specific copies of histone H2A suggest that such duplications may underlie the phenotypic plasticity required for reproductive mode switch in monogononts. We further identified differential expression of genes involved in the formation of resting eggs, a process linked exclusively to sex in this species. Finally, we identified transcripts from the bdelloid rotifer Adineta ricciae that have significant sequence similarity to genes with higher expression in CP strains of B. calyciflorus. Our analysis of global gene expression differences between facultatively sexual and exclusively asexual populations of B. calyciflorus provides insights into the molecular nature of sexual reproduction in rotifers. Furthermore, our results offer insight into the evolution of obligate asexuality in bdelloid rotifers and provide indicators important for the use of monogononts as a model system for investigating the evolution of sexual reproduction.


Cyclosporine and FK506 differentially regulate the sarcolemmal Na(+)-K(+) pump.
Cyclosporine A (CsA) and FK506, important immunosuppressants, have been shown to inhibit the enzymatic equivalent of the Na(+)-K(+) pump (Na(+), K(+)-ATPase) in renal tissue. A similar effect in the heart may contribute to the adverse effects of these agents that include calcification, contractile dysfunction, and altered calcium handling. However, inhibition of the pump has not been demonstrated in cardiac myocytes. We isolated single ventricular myocytes from control rabbits and from rabbits administered CsA or FK506 for 1 week. Na(+)-K(+) pump current (I(p)) was measured using the whole-cell patch-clamp technique. When patch pipettes contained Na(+) in a concentration ([Na](pip)) near physiological intracellular levels mean I(p) of cardiac myocytes from rabbits with serum CsA levels within the therapeutic range was significantly lower than mean I(p) of cardiac myocytes from controls. Treatment had no effect on I(p) measured using a [Na](pip) expected to nearly saturate intracellular binding sites. The CsA-induced inhibition of I(p) was dependent on the K(+) concentration in pipette solutions. Mean I(p) in myocytes from rabbits with serum levels of FK506 within the therapeutic range was similar to mean I(p) in myocytes from controls, whereas FK506 in a dose inducing serum levels severalfold above the therapeutic range caused significant pump inhibition. Using ion-sensitive microelectrodes we showed the intracellular Na(+) activity in papillary muscles isolated from rabbits treated with CsA was significantly higher than in papillary muscles from control rabbits, indicating that CsA causes pump inhibition in intact myocytes with a physiological intracellular milieu.


Acoustothermal Atomization of Water Nanofilms.
We report nonequilibrium molecular simulations of the vibration-induced heating of nanoscale-thick water layers on a metal substrate. In addition to experimentally confirmed acoustothermal evaporation, we observe hitherto unmapped nucleate and film boiling regimes, accompanied by the generation of unprecedented heat fluxes [∼O(10^{9}) W/m^{2}]. We develop a universal scaling parameter to classify the heat-transfer regimes and to predict the thickness of the residual nonevaporating liquid layer. The results find broad application to systems involving drying, coatings, and sprays.


High-dose chemotherapy followed by autologous hematopoietic stem cell transplantation for adult histiocytic disorders with central nervous system involvement.
We postulated that high-dose chemotherapy (HDC) followed by peripheral autologous hematopoietic stem cell transplantation might help to control refractory central nervous system (CNS) histiocytic disorders. Six patients with histiocytic CNS involvement were treated in this way. Two patients achieved non-active disease status, although one relapsed at 84 months. Two patients had regressive disease, one of whom progressed at 21 months. One patient had progressive disease at 14 months. One patient had extra-CNS progression but CNS regression. After a median follow-up of 22.4 months, only one of the six patients still has non-active disease. Treatment was effective on craniofacial and space-occupying brainstem lesions, and was ineffective on neurodegenerative lesions.


Non-immunological hydrops fetalis. a case report.
A case of hydrops fetalis which was not due to isoimmunization is presented. The condition was diagnosed antenatally by means of ultrasonography and the infant was delivered at 32 weeks' gestation. He required intensive care, but survived and is well at 18 months of age. The causation, diagnosis and management of this problem are discussed.


Negative tunneling magnetoresistance by canted magnetization in MgO/NiO tunnel barriers.
The influence of the insertion of an ultrathin NiO layer between the MgO barrier and the ferromagnetic electrodes in magnetic tunnel junctions has been investigated from measurements of the tunneling magnetoresistance and via x-ray magnetic circular dichroism (XMCD). The magnetoresistance shows a high asymmetry with respect to bias voltage, giving rise to a negative value of up to -16% at 2.8 K. We attribute this effect to the formation of noncollinear spin structures at the interface of the NiO layer as inferred from XMCD measurements. The magnetic moments of the interface Ni atoms tilt from their easy axis due to exchange coupling with the neighboring ferromagnetic electrode, and the tilting angle decreases with increasing NiO thickness. The experimental observations are further supported by noncollinear spin density functional calculations.


Working memory predictors of written mathematics in 7- to 8-year-old children.
There is extensive evidence for the involvement of working memory in mathematical attainment. This study aims to identify the relative contributions of verbal, spatial-simultaneous, and spatial-sequential working memory measures in written mathematics. Year 3 children (7-8 years of age, n = 214) in the United Kingdom were administered a battery of working memory tasks alongside a standardised test of mathematics. Confirmatory factor analyses and variance partitioning were then performed on the data to identify the unique variance accounted for by verbal, spatial-simultaneous, and spatial-sequential measures. Results revealed the largest individual contribution was that of verbal working memory, followed by spatial-simultaneous factors. This suggests the components of working memory underpinning mathematical performance at this age are those concerning verbal-numeric and spatial-simultaneous working memory. Implications for educators and further research are discussed.


The World Trade Center Health Program: an introduction to best practices.
More than 20 years have elapsed since the September 11, 2001 (9/11) terrorist attacks on the World Trade Center (WTC), Pentagon and at Shanksville, PA. Many persons continue to suffer a variety of physical and mental health conditions following their exposures to a mixture of incompletely characterized toxicants and psychological stressors at the terrorist attack sites. Primary care and specialized clinicians should ask patients who may have been present at any of the 9/11 sites about their 9/11 exposures, especially patients with cancer, respiratory symptoms, chronic rhinosinusitis, gastroesophageal reflux disease, psychiatric symptoms, and substance use disorders. Clinicians, especially those in the NY metropolitan area, should know how to evaluate, diagnose, and treat patients with conditions that could be associated with exposure to the 9/11 attacks and its aftermath. As such, this issue of Archives contains a series of updates to clinical best practices relevant to medical conditions whose treatment is covered by the WTC Health Program. This first paper in the 14-part series describes the purpose of this series, defines the WTC Health Program and its beneficiaries, and explains how relevant Clinical Practice Guidelines were identified. This paper also reminds readers that because physical and mental health conditions are often intertwined, a coordinated approach to care usually works best and referral to health centers affiliated with the WTC Health Program may be necessary, since all such Centers offer multidisciplinary care.


Comparison of ceftazidime concentrations in bile and serum.
Biliary excretion of ceftazidime, a new broad-spectrum cephalosporin, was studied in two groups of patients after administration of a 2-g dose intravenously. Group A included 10 patients in whom ceftazidime levels in bile were measured during cholecystectomy. Group B included 10 patients with indwelling biliary tubes in whom ceftazidime levels in bile and serum were simultaneously measured at 0.5, 1, 2, 4, 6, and 8 h after administration of the drug. Although ceftazidime levels were variable, they exceeded the minimal inhibitory concentrations of most biliary tract pathogens in both groups.


The inhibin B response in male rats treated with two drug candidates.
Serum Inhibin B was measured in two studies of known testis-toxic drug candidates. Study 1 was for a compound for Hepatitis C, and utilized a 10-week dosing period, followed by mating and necropsy of half of each group, and then a 12-week recovery period for the remaining animals. At the postmating necropsy, 6 of 15 high-dose males had testis lesions; Inhibin B was significantly reduced in all animals in that group. The mid-dose group had no lesions but significantly reduced serum Inhibin B. At recovery, 9 of 15 high-dose males showed damage in testes; serum Inhibin B levels were not different from controls. Inhibin B appeared to both overreport and underreport testis damage in Study 1. Study 2 was an acute pathogenesis study for an antibacterial compound, using control and two dose levels and multiple time points (days 5, 8, 15, 22, and then untreated until day 71). At each time point blood was sampled from all remaining rats and five/group were killed for histologic evaluation. The low-dose group had minimal to moderate lesions, while serum Inhibin B was never changed. The high-dose animals progressed quickly from minimal lesions to being broadly and moderately affected; serum Inhibin B levels were reduced at days 8 and 15 only. In Study 2, Inhibin B appeared less sensitive than histology, except at the extremes of testis damage, when Inhibin B was routinely low. We conclude that in these two studies there was a poor correlation between changes in serum levels of Inhibin B and testis histopathology.


Linearization and state estimation of unknown discrete-time nonlinear dynamic systems using recurrent neurofuzzy networks.
Model-based methods for the state estimation and control of linear systems have been well developed and widely applied. In practice, the underlying systems are often unknown and nonlinear. Therefore, data based model identification and associated linearization techniques are very important. Local linearization and feedback linearization have drawn considerable attention in recent years. In this paper, linearization techniques using neural networks are reviewed, together with theoretical difficulties associated with the application of feedback linearization. A recurrent neurofuzzy network with an analysis of variance (ANOVA) decomposition structure and its learning algorithm are proposed for linearizing unknown discrete-time nonlinear dynamic systems. It can be viewed as a method for approximate feedback linearization, as such it enlarges the class of nonlinear systems that can be feedback linearized using neural networks. Applications of this new method to state estimation are investigated with realistic simulation examples, which shows that the new method has useful practical properties such as model parametric parsimony and learning convergence, and is effective in dealing with complex unknown nonlinear systems.


Carbonic anhydrase inhibitors: X-ray crystallographic structure of the adduct of human isozyme II with a bis-sulfonamide-two heads are better than one?
The X-ray crystal structure for the adduct of human carbonic anhydrase II (hCA II) with 4-(4-sulfamoylphenylcarboxamidoethyl)benzenesulfonamide, a topically acting antiglaucoma sulfonamide has been resolved at a resolution of 1.8 A. Its binding to the enzyme is similar with that of other sulfonamides, considering the interactions of the sulfonamide zinc anchoring group, but differs considerably when the organic part of the inhibitor is analyzed. This part of the inhibitor interacts only within the hydrophobic half of the CA active site, leaving the hydrophilic half able to accomodate several water molecules not present in the uncomplexed enzyme. Furthermore, the second head (sulfonamide moiety) participates in two strong hydrogen bonds with amino acid residues (Gly 132 and Gln 136) situated on the rim of the entrance to the active site cleft. Thus, the answer to the question in the title of this paper is that two heads are better than one, since the two sulfamoyl moieties of the inhibitor allow its proper orientation within the active site, with only one head binding in ionized form to the zinc ion, the organic part lying within the hydrophobic half of the active site, and the terminal, carboxamido containing phenylsulfamoyl head participating in strong hydrogen bonds with amino acid residues located at the entrance of it. All these findings are important for the design of better carboxamido CA inhibitors with applications in clinical medicine.


The formic acid-nitric acid complex: microwave spectrum, structure, and proton transfer.
Rotational spectra are reported for seven isotopologues of the complex HCOOH-HNO3 in a supersonic jet. The system is planar and bound by a pair of hydrogen bonds, much like the more widely studied carboxylic acid dimers. Double proton exchange interconverts the system between a pair of equivalent structures, as revealed by a splitting of the a-type spectrum that disappears when one of the hydrogen bonding protons is replaced by deuterium. The observation of relative intensities that are consistent with nuclear spin statistics in a symmetric and antisymmetric pair of tunneling states provides additional evidence for such a motion. The observed splittings in the pure rotational spectrum are 1-2 orders of magnitude smaller than those recently reported in the pure rotational spectra of several related carboxylic acid dimers. This is a curious difference, although we note that because the observed spectra do not cross the tunneling doublet, the splittings are a measure of the difference in effective rotational constants for the two states, not the tunneling frequency itself. The observed rotational constants have been used to determine an accurate vibrationally averaged structure for the complex. The two hydrogen bond lengths, 1.686(17) Å and 1.813(10) Å for the hydrogen bonds involving the HNO3 and HCOOH protons, respectively, differ by 0.127(27) Å. Likewise, the associated oxygen-oxygen distances determined for the parent species, 2.631 and 2.794 Å, differ by 0.163 Å. These results suggest that the double proton transfer is necessarily accompanied by substantial motion of the heavy atom frame, and thus this system, in principle, provides an excellent prototype for multidimensional tunneling processes. Ab initio calculations of the binding energy and the barrier height are presented. Excellent agreement between the calculated equilibrium structure and the experimental, vibrationally averaged structure suggests that the vibrational wave function is not highly delocalized in the region between the equivalent potential wells. (14)N nuclear quadrupole hyperfine structure is interpreted in terms of the degree to which the HNO3 releases its proton in either of the equivalent potential energy minima.


Prediction of the in vitro permeability determined in Caco-2 cells by using artificial neural networks.
Caco-2 cells are currently the most used in vitro tool for prediction of the potential oral absorption of new drugs. The existence of computational models based on this data may potentiate the early selection process of new drugs, but the current models are based on a limited number of cases or on a reduced molecular space. We present an artificial neural network based only on calculated molecular descriptors for modelling 296 in vitro Caco-2 apparent permeability (P(app)) drug values collected in the literature using also a pruning procedure for reducing the descriptors space. LogP(app) values were divided into a training group of 192 drugs for network optimization and a testing group of another 59 drugs for early stop and internal validation resulting in correlations of 0.843 and 0.702 and RMSE of 0.546 and 0.791 for the training and testing group, respectively. External validation was made with an additional group of 45 drugs with a correlation of 0.774 and RMSE of 0.601. The selected molecular descriptors encode information related to the lipophilicity, electronegativity, size, shape and flexibility characteristics of the molecules, which are related to drug absorption. This model may be a valuable tool for prediction and simulation in the drug development process, as it allows the in silico estimation of the in vitro Caco-2 apparent permeability.


Reduced integration and improved segregation of functional brain networks in Alzheimer's disease.
Emerging evidence shows that cognitive deficits in Alzheimer's disease (AD) are associated with disruptions in brain functional connectivity. Thus, the identification of alterations in AD functional networks has become a topic of increasing interest. However, to what extent AD induces disruption of the balance of local and global information processing in the human brain remains elusive. The main objective of this study is to explore the dynamic topological changes of AD networks in terms of brain network segregation and integration. We used electroencephalography (EEG) data recorded from 20 participants (10 AD patients and 10 healthy controls) during resting state. Functional brain networks were reconstructed using EEG source connectivity computed in different frequency bands. Graph theoretical analyses were performed assess differences between both groups. Results revealed that AD networks, compared to networks of age-matched healthy controls, are characterized by lower global information processing (integration) and higher local information processing (segregation). Results showed also significant correlation between the alterations in the AD patients' functional brain networks and their cognitive scores. These findings may contribute to the development of EEG network-based test that could strengthen results obtained from currently-used neurophysiological tests in neurodegenerative diseases.


Efficacy and Safety of Low-Dose Liposomal Amphotericin B in Adult Patients Undergoing Unrelated Cord Blood Transplantation.
Liposomal amphotericin B (L-AMB) is widely used for empirical or preemptive therapy and treatment of invasive fungal infections after cord blood transplantation (CBT). We retrospectively examined the efficacy and safety of low-dose L-AMB in 48 adult patients who underwent CBT between 2006 and 2017 in our institute. Within the entire cohort, 42 patients (88%) received L-AMB as empirical or preemptive therapy. The median daily dose of L-AMB and the median cumulative dose of L-AMB were 1.20 mg/kg/day (range, 0.62 to 2.60 mg/kg/day) and 30.6 mg/kg (range, 0.7 to 241.5 mg/kg), respectively. The median duration of L-AMB administration was 21.5 days (range, 1 to 313 days). A documented breakthrough fungal infection occurred in 1 patient during L-AMB treatment, and 43 patients (90%) survived for at least 7 days after the end of L-AMB treatment. Grade 3 or higher hypokalemia and hepatotoxicity were frequently observed during L-AMB treatment. However, no patient developed an increase in serum creatinine levels of grade 3 or higher. In univariate analyses using a logistic regression model, a duration of L-AMB treatment of more than 21 days and a cumulative dose of L-AMB of more than 30 mg/kg were significantly associated with nephrotoxicity and grade 3 hypokalemia. These data suggest that low-dose L-AMB may be safe and effective in adult patients undergoing CBT.


Broader visual orientation tuning in patients with schizophrenia.
Reduced gamma-aminobutyric acid (GABA) levels in cerebral cortex are thought to contribute to information processing deficits in patients with schizophrenia (SZ), and we have previously reported lower in vivo GABA levels in the visual cortex of patients with SZ. GABA-mediated inhibition plays a role in sharpening orientation tuning of visual cortical neurons. Therefore, we predicted that tuning for visual stimulus orientation would be wider in SZ. We measured orientation tuning with a psychophysical procedure in which subjects performed a target detection task of a low-contrast oriented grating, following adaptation to a high-contrast grating. Contrast detection thresholds were determined for a range of adapter-target orientation offsets. For both SZ and healthy controls, contrast thresholds decreased as orientation offset increased, suggesting that this tuning curve reflects the selectivity of visual cortical neurons for stimulus orientation. After accounting for generalized deficits in task performance in SZ, there was no difference between patients and controls for detection of target stimuli having either the same orientation as the adapter or orientations far from the adapter. However, patients' thresholds were significantly higher for intermediate adapter-target offsets. In addition, the mean width parameter of a Gaussian fit to the psychophysical orientation tuning curves was significantly larger for the patient group. We also present preliminary data relating visual cortical GABA levels, as measured with magnetic resonance spectroscopy, and orientation tuning width. These results suggest that our finding of broader orientation tuning in SZ may be due to diminished visual cortical GABA levels.


Various glycolysis inhibitor-containing tubes for glucose measurement cannot be used interchangeably due to clinically unacceptable biases between them.
The aim of our study was to determine the difference between glucose concentration measured 30 min after venipuncture in ice-chilled heparin plasma sample and all currently available citrate buffer-containing tubes (Greiner Glucomedics, Greiner FC Mix and Sarstedt GlucoEXACT) and still widely used sodium fluoride/potassium oxalate (NaF/Kox) tubes from Greiner. Blood was collected from 20 healthy volunteers and 20 patients with diabetes into LiH, NaF/KOx, Glucomedics, FC mix and GlucoEXACT tubes. Glucose was measured within 30 min from blood sampling in duplicate on the Architect c8000 analyzer. Mean biases between all tube types were calculated and compared to the recommended criteria (1.95%). Additionally, glucose concentrations measured in all five tube types were compared using the Friedman test. In the entire studied population, glucose concentrations measured in Glucomedics, FC mix and GlucoEXACT were higher (7.3%, 3.2% and 2.0%, respectively) than in the ice-chilled LiH tubes. When all glycolysis inhibitor-containing tubes were compared, Glucomedics tubes significantly differed from GlucoEXACT and FC mix tubes (biases -4.9% and 4.0%, respectively). In addition, there was a significant difference between the NaF/KOx tube and Glucomedics, as well as FC mix tubes (biases 7.1% and 3.0%, respectively). Glucose concentrations measured in recommended ice-chilled lithium heparin- and citrate buffer-containing tubes are not comparable. Significant biases exist between various glycolysis inhibitor-containing tubes; therefore, they cannot be used interchangeably.


1,3-Alternate Calix[4]arene Functionalized With Pyrazole and Triazole Ligands as a Highly Selective Fluorescent Sensor for Hg2+ and Ag+ Ions.
We report here the synthesis of a 1,3-alternate calix[4]arene 8, with bis-pyrazolylmethylpyrenes on the one end and bis-triazolylmethylphenyls on the other end, as a homoditropic fluorescent sensor for both Hg2+ and Ag+ ions. Calix[4]arene 3, with lower-rim bis-pyrazolylmethylpyrenes in cone conformation, was also synthesized as a control compound. UV-Vis and fluorescence spectra were used for metal ions screening, and we found that both ligands 8 and 3 showed strong excimer emission of pyrenes when they are as a free ligand in CHCl3/MeOH (v/v, 3:1) solution; however, they both showed a high selectivity toward Hg2+ and Ag+ ions with strong fluorescence quenching and yet with different binding ratios. The fluorescence of ligand 8 was strongly quenched by Hg2+ but was only partially quenched by Ag+ ions; however, the fluorescence of ligand 3 was strongly quenched by Hg2+, Ag+, and Cu2+ ions. Job plot experiments showed that ligand 8 formed a 1:2 complex with both Hg2+ and Ag+ ions; ligand 3 formed a 1:1 complex with Hg2+, but it formed a 2:3 complex with Ag+. The binding constant of ligand 3 with Hg2+ and Ag+ ions was determined by the Benesi-Hildebrand plot of UV-vis titration experiments to be 2.99 × 103 and 3.83 × 103 M-1, respectively, while the association constant of ligand 8 with Hg2+ and Ag+ was determined by Hill plot to be 1.46 × 1012 and 9.24 × 1011 M-2, respectively. Ligand 8 forms a strong complex with either two Hg2+ or two Ag+ ions using both the upper and lower rims of the 1,3-alternate calix[4]arene as the binding pockets; hence, it represents one of the highly selective fluorescent sensors for the homoditropic sensing of Hg2+ and Ag+ ions.


A mesocosm study of the effects of wet-dry cycles on nutrient release from constructed wetlands in agricultural landscapes.
Given the projection that wet-dry periods will be more frequent in the US Midwest, a study was conducted to understand the impact of these hydro-climatic alterations on nutrient dynamics in wetlands constructed on former croplands in the region. Soil cores were collected from two constructed wetlands and a wooded riparian area (surface: 0-20 cm; subsurface: 40-60 cm) downslope from an agricultural field. Cores were either kept moist or subjected to a 5-week drying treatment, after which all cores were flooded for 36 days. Initial nitrate flux was significantly (p < 0.001) higher in the dry than in the moist treatment (44.5 vs. 1.9 mg N m(-2) per day), likely due to mineralization of organic matter. The NO3(-) released was rapidly denitrified (N2O flux: 18.9 mg N m(-2) per day), except in the subsurface soil cores in which processing of available N (N2O flux: 0.33 mg N m(-2) per day) was limited by low microbial activity (4 times lower CO2 production rate). The dry treatment also resulted in significantly (p < 0.01) higher inorganic P (Pi) flux (3.1 versus 1 mg P m(-2) per day in moist cores), with water-extractable soil P being the best predictor (r(2): 0.93, p < 0.03) of that flux. Despite a decline in redox potential (as low as -36.4 mv) and progressive increase in pore-water dissolved Fe, no relationship between floodwater Pi and dissolved Fe was observed, suggesting either limited contribution of reductive dissolution to Pi dynamics or rapid adsorption of the Pi released within the cores. Compared to the moist cores, geochemical modeling showed a consistent shift toward greater solubility of the calcium-phosphate minerals controlling pore-water Pi concentration in the dry treatment cores. These results suggest that dissolution of Ca-phosphate minerals could be a key factor controlling Pi mobility in constructed wetlands subjected to wet-dry cycles.


Effects of sibutramine treatment in obese adolescents: a randomized trial.
Increased prevalence of adolescent obesity requires effective treatment options beyond behavior therapy. To see whether sibutramine reduced weight more than placebo in obese adolescents who were receiving a behavior therapy program. 12-month, 3:1 randomized, double-blind trial conducted from July 2000 to February 2002. 33 U.S. outpatient clinics. 498 participants 12 to 16 years of age with a body mass index (BMI) that was at least 2 units more than the U.S. weighted mean of the 95th percentile based on age and sex, to the upper limit of 44 kg/m2. Site-specific behavior therapy plus 10 mg of sibutramine or placebo. Blinded study medication dose was uptitrated to 15 mg or placebo at month 6 if initial BMI was not reduced by 10%. Body mass index, waist circumference, body weight, fasting lipid and glycemic variables, safety, and tolerability. Seventy-six percent of patients in the sibutramine group and 62% of patients in the placebo group completed the study. The estimated mean treatment group difference at month 12 (linear mixed-effects model) favored sibutramine for change from baseline in BMI (-2.9 kg/m2 [95% CI, -3.5 to -2.2 kg/m2]) and body weight (-8.4 kg [CI, -9.7 to -7.2 kg]) (P < 0.001 for both). The sibutramine group had greater improvements in triglyceride levels, high-density lipoprotein cholesterol levels, insulin levels, and insulin sensitivity (P < or = 0.001 for all). The rate of tachycardia was greater with sibutramine vs. placebo (12.5% vs. 6.2%; difference, 6.3 percentage points [CI, 1.0 to 11.7 percentage points]) but did not lead to increased withdrawal (2.4% vs. 1.5%; difference, 0.9 percentage point [CI, -1.7 to 3.5 percentage points]). The 1-year study duration precluded assessment of long-term weight maintenance and putative health benefits and harms, and 24% and 38% of the sibutramine and placebo groups, respectively, did not complete follow-up. Sibutramine added to a behavior therapy program reduced BMI and body weight more than placebo and improved the profile of several metabolic risk factors in obese adolescents.


Impact of Bifidobacterium longum1714® on maternal cytokine response in peripheral blood mononuclear cells.
The maternal immune system is implicated in adverse pregnancy outcomes. Manipulation of maternal immune response by probiotics holds potential to reduce pregnancy complications. The MicrobeMom2 study investigates the impact of probiotic supplementation on maternal immune responses to pathogen associated molecular patterns (PAMPs) in peripheral blood mononuclear cells (PBMCs) during pregnancy. This double-blinded randomised-controlled trial involved oral supplementation of Bifidobacterium longum subsp. longum 1714® (B. longum 1714; daily ingestion of a minimum of 1x109 colony forming units) or placebo from 16 to 20-weeks' gestation until delivery in healthy pregnant women. The primary outcome was a change in IL-10 production, after stimulation with Lipopolysaccharide (LPS) or anti-CD3/28/2, in PBMCs isolated from blood samples taken at baseline (11-15 weeks' gestation) and late pregnancy (28-32 weeks' gestation) after 48 h incubation. 68 subjects were needed (34ineachgroup) for 80 % power at an alpha significance of 0.05 to detect differences in IL10. 72 women (mean ± SD age 33.17 ± 4.53 years and median (25th, 75th centile) body mass index 24.93 (21.93, 27.57 kg/m2)) were recruited with primary outcome data. Using LPS, late pregnancy fold change in IL-10 in PBMCs after 48 h incubation was median (25th, 75th centile) 88.45 (4.88, 488.78) in the intervention, 24.18 (6.36, 141.17) in the control group, p = 0.183. Using anti-CD3/28/2, values were 189.69 (425.96, 866.57),148.74 (31.67, 887.03) in intervention and control groups, respectively, p = 0.506. No significant differences were observed between the two groups. Maternal antenatal supplementation with B. longum 1714 did not alter cytokine production by maternal PBMCs in response to PAMPs or anti-CD3/28/2. ISRCTN registry ISRCTN43013285.


Differential regulation of hippocampal progenitor proliferation by opioid receptor antagonists in running and non-running spontaneously hypertensive rats.
Voluntary running in mice and forced treadmill running in rats have been shown to increase the amount of proliferating cells in the hippocampus. Little is known as yet about the mechanisms involved in these processes. It is well known that the endogenous opioid system is affected during running and other forms of physical exercise. In this study, we evaluated the involvement of the endogenous opioids in the regulation of hippocampal proliferation in non-running and voluntary running rats. Nine days of wheel running was compared with non-running in spontaneously hypertensive rats (SHR), a rat strain known to run voluntarily. On the last 2 days of the experimental period all rats received two daily injections of the opioid receptor antagonists naltrexone or naltrindole together with injections of bromodeoxyuridine to label dividing cells. Brain sections from the running rats showed approximately a five-fold increase in newly generated cells in the hippocampus, and this increase was partly reduced by naltrexone but not by naltrindole. By contrast, both naltrexone and naltrindole increased hippocampal proliferation in non-running rats. In non-running rats the administration of naltrexone decreased corticosterone levels and adrenal gland weights, whereas no significant effects on these parameters could be detected for naltrindole. However, adrenal gland weights were increased in naltrexone- but not in naltrindole-administered running rats. In addition, in voluntary running rats there was a three-fold increase in the hippocampal levels of Met-enkephalin-Arg-Phe compared with non-runners, indicating an increase in opioid activity in the hippocampus during running. These data suggest an involvement of endogenous opioids in the regulation of hippocampal proliferation in non-running rats, probably through hypothalamic-pituitary-adrenal axis modulation. During voluntary running in SHR naltrexone altered hippocampal proliferation via as yet unknown mechanisms.


Selective complement C1s deficiency caused by homozygous four-base deletion in the C1s gene.
The complement system plays an important role in defense mechanisms by promoting the adherence of microorganisms to phagocytic cells and lysis of foreign organisms. Deficiencies of the first complement components, C1r/C1s, often cause systemic lupus erythematosus-like syndromes and severe pyogenic infections. Up to now no genetic analysis of the C1r/C1s deficiencies has been carried out. In the present work, we report the first genetic analysis of selective C1s deficiency, the patient having a normal amount of C1r. C1s RNA with a normal size was detected in patient's subcutaneous fibroblasts (YKF) by RNA blot analysis and RT-PCR. The amount of C1s RNA was approximately one-tenth of the RNA from the human chondrosarcoma cell line, HCS2/8. In contrast, the levels of C1r and beta-actin RNA of YKF were similar to that of HCS2/8. Sequence analysis of C1s cDNA revealed a deletion at nucleotides 1087-1090 (TTTG), creating a stop codon (TGA) at position 94 downstream of the mutation site. Direct sequencing of the gene between the primers designed on intron 9 and exon 10 indicated the presence of the deletion on exon 10 of the gene. Quantitative Southern blot hybridization suggested the mutation was homozygous. The 4-bp deletion on exon 10 was also found in the patient's heterozygous mother who had normal hemolytic activity.


The Unique Lifestyle of Crohn's Disease-Associated Adherent-Invasive Escherichia coli.
Escherichia coli is one of the most genetically and phenotypically diverse species of bacteria. This remarkable diversity produces a plethora of clinical outcomes following infection and has informed much of what we currently know about host-pathogen interactions for a wide range of bacteria-host relationships. In studying the role of microbes in disease, adherent-invasive E. coli (AIEC) has emerged as having a strong association with Crohn's disease (CD). Thus, there has been an equally strong effort to uncover the root origins of AIEC, to appreciate how AIEC differs from other well-known pathogenic E. coli variants, and to understand its connection to disease. Emerging from a growing body of research on AIEC is the understanding that AIEC itself is remarkably diverse, both in phylogenetic origins, genetic makeup, and behavior in the host setting. Here, we describe the unique lifestyle of CD-associated AIEC and review recent research that is uncovering the inextricable link between AIEC and its host in the context of CD.


Characterization of lymphocyte function-associated antigen 1 (LFA-1)-deficient T cell lines: the alphaL and beta2 subunits are interdependent for cell surface expression.
The leukocyte, or beta2, integrins are heterodimeric cell surface molecules that share a common beta subunit, and have unique alpha subunits. LFA-1 is the predominant beta2 integrin on lymphocytes, and plays an important role in many lymphocyte functions; however, most studies of the cytoplasmic regions of LFA-1 have been performed in transfected epithelial cells, such as COS, in part because no lymphoid cell lines deficient in the LFA-1 alpha subunit have been described. To address structure-function studies of beta2 integrins in relevant cell types, two T lymphoblastoid cell clones that completely lack cell surface LFA-1, J-(beta2).7 and SK-(beta2).7, derived from Jurkat and SKW3, respectively, were prepared by chemical mutagenesis and selection. Biosynthetic labeling and immunoprecipitation showed that the J-(beta2).7 clone did not translate any LFA-1 alpha subunit protein, while the SK-(beta2).7 cells did not synthesize any beta2 subunit protein. Northern blot analysis of poly(A+) RNA from these cells revealed an absence of the corresponding mRNA in each case. By transfection analysis, only the alpha subunit reconstituted LFA-1 expression in the J-(beta2).7 cells, while only the beta subunit restored cell surface LFA-1 expression in the SK-(beta2).7 cells. Functional studies with the parental cell lines, the J-(beta2).7 and SK-(beta2).7 cells, and the transfectants showed that all binding of Jurkat and SKW3 cells to purified ICAM-1 is mediated by LFA-1, and the reconstituted LFA-1 expressed by the J-(beta2).7 and SK-(beta2).7 transfected cells is regulated normally.


Personal identity verification for regional emergency workers.
South Central Texas emergency planners have organised a regional Personal Identity Verification (PIV) project for hospital and emergency workers. This paper describes the development and administrative challenges in designing and implementing an electronic PIV system across multiple organisations in a 22-county region.


Influence of BaTiO3 ferroelectric orientation for electro-optic modulation on silicon.
The influence of BaTiO(3) ferroelectric domain orientations for high efficiency electro-optic modulation has been thoroughly analyzed. The Mach-Zehnder modulator structure is based on a CMOS compatible silicon/BaTiO(3)/silicon slot waveguide that supports both TE and TM polarizations whereas the Pockels effect is exploited by the application of a horizontal electric field with lateral electrodes placed on top of the BaTiO(3) layer. The influence of the waveguide parameters has been optimized for each configuration and the lowest V(π) voltage combined with low losses has been determined. A V(π)L as low as 0.27 V·cm has been obtained for a-axis oriented BaTiO(3) and TE polarization by rotating the waveguide structure to an optimum angle.


Development and verification of fingerprinting probes for Candida glabrata.
Since Candida glabrata has emerged as one of the most common Candida pathogens, DNA probes have been developed which fulfil the requirements for effective fingerprinting. Using a screen for complex genomic fragments containing moderately repetitive sequences, seven DNA probes were cloned which generate complex Southern blot hybridization patterns with EcoRI-digested C. glabrata DNA. All of the probes are species-specific and the majority cross-hybridize to varying degrees. The capacity of two of the probes, Cg6 and Cg12, to measure genetic distance between independent isolates is verified by comparing clustering in dendrograms based on similarity coefficients computed between all pairs of 39 independent isolates fingerprinted with Cg6, Cg12 and randomly amplified polymorphic DNA. The capacity of the probes Cg6 and Cg12 to assess microevolution in clonal populations of infecting C. glabrata over time is also demonstrated. These probes can now be used in large computer-assisted epidemiological studies.


α-Synuclein RT-QuIC assay in cerebrospinal fluid of patients with dementia with Lewy bodies.
We applied RT-QuIC assay to detect α-synuclein aggregates in cerebrospinal fluid (CSF) of patients with suspected Creutzfeldt-Jakob disease who had a neuropathological diagnosis of dementia with Lewy bodies (DLB) (n = 7), other neurodegenerative diseases with α-synuclein mixed pathology (n = 20), or without Lewy-related pathology (n = 49). The test had a sensitivity of 92.9% and specificity of 95.9% in distinguishing α-synucleinopathies from non-α-synucleinopathies. When performed in the CSF of patients with DLB (n = 36), RT-QuIC was positive in 17/20 with probable DLB, 0/6 with possible DLB, and 0/10 with Alzheimer disease. These results indicate that RT-QuIC for α-synuclein is an accurate test for DLB diagnosis.


Functional changes and motility characteristics of Japanese Black bull spermatozoa separated by Percoll.
Spermatozoa from two Japanese Black bulls (Bull-ATF and Bull-KTG) were separated by centrifugation at 700 x g for 15min in modified TALP with or without 45-90% Percoll. Control washed spermatozoa and those collected from the bottom of 45 and 90% Percoll fractions were examined for viability and membrane integrity (using Hoechst bis-benzimide 33258 or propidium iodide and 6-carboxyfluorescein diacetate (PI-CFDA)), acrosomal status (using fluorescence isothiocyanate (FITC) conjugated Pisum Sativum agglutinin (PSA) and Peanut agglutinin (PNA), Naphthol Yellow S and Erythrosin B (NE) or triple staining (TS)), capacitation status (using chlortetracycline (CTC)), motility characteristics (using a computer-assisted sperm motion analysis system (CASA)) and for in vitro fertility. Percoll-separated spermatozoa showed greater viability and membrane integrity than controls, as determined by supravital staining. Differences were observed in the results regarding viability and acrosomal status of spermatozoa among sperm staining methods. Bull-ATF, which showed significantly greater in vitro fertility than Bull-KTG (P<0.05), showed a significantly higher rate of CTC-B-pattern (capacitated) spermatozoa (P<0.01) than Bull-KTG. The motility characteristics of control washed spermatozoa and those separated by 45-90% Percoll were analyzed by CASA. More motile and progressively motile spermatozoa were observed in the fraction at the bottom of the 90% Percoll solution than in the 45% Percoll fraction or in controls (P<0.01). Moreover, the spermatozoa of Bull-KTG, which showed lower in vitro fertility than Bull-ATF, did not show significant differences in motility from those of Bull-ATF. These results provided basic information about Japanese Black bull spermatozoa, and suggested that spermatozoa with greater motility and viability can be obtained by Percoll separation than without separation. However, Percoll separation did not enhance their in vitro fertility.