Progress and Challenges in the Biology of FNDC5 and Irisin.
In 2002, a transmembrane protein-now known as FNDC5-was discovered and shown to be expressed in skeletal muscle, heart, and brain. It was virtually ignored for 10 years, until a study in 2012 proposed that, in response to exercise, the ectodomain of skeletal muscle FNDC5 was cleaved, traveled to white adipose tissue, and induced browning. The wasted energy of this browning raised the possibility that this myokine, named irisin, might mediate some beneficial effects of exercise. Since then, more than 1000 papers have been published exploring the roles of irisin. A major interest has been on adipose tissue and metabolism, following up the major proposal from 2012. Many studies correlating plasma irisin levels with physiological conditions have been questioned for using flawed assays for irisin concentration. However, experiments altering irisin levels by injecting recombinant irisin or by gene knockout are more promising. Recent discoveries have suggested potential roles of irisin in bone remodeling and in the brain, with effects potentially related to Alzheimer's disease. We discuss some discrepancies between research groups and the mechanisms that are yet to be determined. Some important questions raised in the initial discovery of irisin, such as the role of the mutant start codon of human FNDC5 and the mechanism of ectodomain cleavage, remain to be answered. Apart from these specific questions, a promising new tool has been developed-mice with a global or tissue-specific knockout of FNDC5. In this review, we critically examine the current knowledge and delineate potential solutions to resolve existing ambiguities.


Distinct Neural Components of Visually Guided Grasping during Planning and Execution.
Selecting suitable grasps on three-dimensional objects is a challenging visuomotor computation, which involves combining information about an object (e.g., its shape, size, and mass) with information about the actor's body (e.g., the optimal grasp aperture and hand posture for comfortable manipulation). Here, we used functional magnetic resonance imaging to investigate brain networks associated with these distinct aspects during grasp planning and execution. Human participants of either sex viewed and then executed preselected grasps on L-shaped objects made of wood and/or brass. By leveraging a computational approach that accurately predicts human grasp locations, we selected grasp points that disentangled the role of multiple grasp-relevant factors, that is, grasp axis, grasp size, and object mass. Representational Similarity Analysis revealed that grasp axis was encoded along dorsal-stream regions during grasp planning. Grasp size was first encoded in ventral stream areas during grasp planning then in premotor regions during grasp execution. Object mass was encoded in ventral stream and (pre)motor regions only during grasp execution. Premotor regions further encoded visual predictions of grasp comfort, whereas the ventral stream encoded grasp comfort during execution, suggesting its involvement in haptic evaluation. These shifts in neural representations thus capture the sensorimotor transformations that allow humans to grasp objects.SIGNIFICANCE STATEMENT Grasping requires integrating object properties with constraints on hand and arm postures. Using a computational approach that accurately predicts human grasp locations by combining such constraints, we selected grasps on objects that disentangled the relative contributions of object mass, grasp size, and grasp axis during grasp planning and execution in a neuroimaging study. Our findings reveal a greater role of dorsal-stream visuomotor areas during grasp planning, and, surprisingly, increasing ventral stream engagement during execution. We propose that during planning, visuomotor representations initially encode grasp axis and size. Perceptual representations of object material properties become more relevant instead as the hand approaches the object and motor programs are refined with estimates of the grip forces required to successfully lift the object.


Quantification of anti-drug antibodies against E6011, an anti-fractalkine monoclonal antibody, in monkey and human serum, by an electrochemiluminescence assay.
E6011, a humanized anti-fractalkine monoclonal antibody, is under development for the treatment of various inflammatory diseases, such as rheumatoid arthritis. Therapeutic antibodies may induce production of anti-drug antibodies (ADA) that may deteriorate efficacy and/or enhance immunogenic reaction. It is important to have an ADA assay to understand the characteristics of biotherapeutics under development. A simple and reproducible assay has thus been developed for the determination of ADA against E6011 in monkey and human serum by electrochemiluminescence (ECL) detection. An immune-complex of biotinylated E6011, ADA, and ruthenium-labeled E6011 was attached to avidin-coated wells for ECL signal detection. Screening and confirmatory cutpoints were determined to judge negative or positive ADA. Sensitivity of ADA was 1.61 and 1.34 ng/mL in monkey and human serum, respectively. Accuracy and precision of the assay were within ±20% and 20%, respectively. Drug tolerance of the assay in monkey and human sera was ensured up to 100 and 1000 μg/mL E6011 at the surrogate ADA levels of 1 and 4 μg/mL, respectively. The developed assay was successfully applied to ADA quantification in monkeys and humans in support of immunogenicity assessments.


Design of a problem-based curriculum: a general approach and a case study in the domain of public health.
A general approach to curriculum design in the context of Problem-Based Learning (PBL) is outlined. Ten general 'steps' for problem-based curriculum development are proposed, using the case study methodology to describe the underlying iterative process. Examples are given from the case of the development of a public health professions education curriculum. The process starts with defining the purpose of the curriculum. General objectives are generated in a top-down fashion. The prior knowledge, skills and misconceptions of future students are considered. A preliminary schedule of the curriculum is developed, including sketches of unit blueprints. These are further elaborated. Unit subgoals are related to planned educational activities. Only then, are the learning materials created, with problem writing as the most important aspect. In developing a problem-based curriculum, assessment deserves special attention, because of its influence on the learning process. Finally, educational organization, curriculum management and evaluation procedures are considered.


Glucose 6-Phosphate dehydrogenase deficiency genetic, pathophysiological and therapeutic aspects.
Glucose 6-phosphate dehydrogenase (G6PD) is the first enzyme of the pentose phosphate pathway. It exists in over 250 variants which are divided broadly into five classes on the basis of residual enzyme activity and clinical manifestations. The variants with reduced activity result in G6PD deficiency. This is inherited as an X-linked recessive disorder and occurs at a much higher frequency in the male than in the female. Most G6PD deficient individuals show no clinical abnormality under normal conditions, but acute hemolytic crisis may occur. Several nonhemolytic abnormalities occur as well in G6PD deficients at a higher frequency than in nondeficients. Several nonhemolytic abnormalities occur as well in G6PD deficients at a higher frequency than in nondeficients. The genetic, pathophysiologic, and therapeutic aspects of G6PD deficiency are presented; and the possibility of genetic counselling, care in blood banks, and benefits of education the G6PD deficients are discussed.


Pressure-induced transformation of CH3NH3PbI3: the role of the noble-gas pressure transmitting media.
The photovoltaic perovskite, methylammonium lead triiodide [CH3NH3PbI3 (MAPbI3)], is one of the most efficient materials for solar energy conversion. Various kinds of chemical and physical modifications have been applied to MAPbI3 towards better understanding of the relation between composition, structure, electronic properties and energy conversion efficiency of this material. Pressure is a particularly useful tool, as it can substantially reduce the interatomic spacing in this relatively soft material and cause significant modifications to the electronic structure. Application of high pressure induces changes in the crystal symmetry up to a threshold level above which it leads to amorphization. Here, a detailed structural study of MAPbI3 at high hydrostatic pressures using Ne and Ar as pressure transmitting media is reported. Single-crystal X-ray diffraction experiments with synchrotron radiation at room temperature in the 0-20 GPa pressure range show that atoms of both gaseous media, Ne and Ar, are gradually incorporated into MAPbI3, thus leading to marked structural changes of the material. Specifically, Ne stabilizes the high-pressure phase of NexMAPbI3 and prevents amorphization up to 20 GPa. After releasing the pressure, the crystal has the composition of Ne0.97MAPbI3, which remains stable under ambient conditions. In contrast, above 2.4 GPa, Ar accelerates an irreversible amorphization. The distinct impacts of Ne and Ar are attributed to differences in their chemical reactivity under pressure inside the restricted space between the PbI6 octahedra.


Hemodynamic indices of myocardial dysfunction correlate with dipyridamole thallium-201 SPECT.
Important differences in hemodynamics and tracer kinetics occur with dipyridamole compared to exercise scintigraphy. To better understand the clinical significance of dipyridamole SPECT 201Tl scintigraphy, we examined the relationships between scintigraphy and clinical, and angiographic and hemodynamic variables in patients with CAD. Forty-nine subjects were divided into three study groups. Patients in Groups A (n = 11) and B (n = 20) had a low (<5%) likelihood of CAD. Group A underwent maximal exercise thallium stress testing. Group B underwent thallium dipyridamole scintigraphy. Group C (n = 18) consisted of patients with coronary artery disease who had dipyridamole thallium scintigraphy and cardiac catheterization within 2 wk. Thallium lung-to-myocardial ratio (L/M), left ventricular dilation and perfusion defect site were compared to hemodynamic, clinical and angiographic variables. The Group A L/M ratio of 0.23 +/- 0.05 (mean +/- 1 s.d.) was significantly lower (p < 0.001) compared to the Group B L/M ratio of 0.31 +/- 0.05. In Group C, the UM ratio showed correlation with indices of left ventricular dysfunction including lower resting ejection fraction (p = 0.02, r = 0.83), higher pulmonary capillary wedge pressure (p = 0.01, r = 0.58) and lower cardiac index (p = 0.03, r = 0.54). Left ventricular dilation was associated with hemodynamic changes of ventricular failure including lower resting ejection fraction (p = 0.008, r = 0.88) and higher pulmonary capillary wedge pressure (p = 0.02, r =0.54). Immediate and delayed perfusion defect size showed good correlation with lower resting left ventricular ejection fraction (p = 0.02, r = 0.83, and p = 0.004, r = 0.91, respectively). Lung uptake, left ventricular dilation and perfusion defect size show good correlation to hemodynamic indices of resting left ventricular dysfunction. A combination of these factors may be a better predictor of future cardiac events and prognosis.


Procoagulant activity in patients with combined type 2 diabetes and coronary artery disease: No effects of long-term exercise training.
We investigated the effects of 12-month exercise training on hypercoagulability in patients with combined type 2 diabetes mellitus and coronary artery disease. Associations with severity of disease were further explored. Patients ( n = 131) were randomized to exercise training or a control group. Blood was collected at inclusion and after 12 months. Tissue factor, free and total tissue factor pathway inhibitor, prothrombin fragment 1 + 2 (F1 + 2) and D-dimer were determined by enzyme-linked immunosorbent assay and ex vivo thrombin generation by the calibrated automated thrombogram assay. Tissue factor and ex vivo thrombin generation increased from baseline to 12 months ( p < 0.01, all), with no significant differences in changes between groups. At baseline, free and total tissue factor pathway inhibitor significantly correlated to fasting glucose ( p < 0.01, both) and HbA1c ( p < 0.05, both). In patients with albuminuria ( n = 34), these correlations were strengthened, and elevated levels of D-dimer, free and total tissue factor pathway inhibitor ( p < 0.01, all) and decreased ex vivo thrombin generation ( p < 0.05, all) were observed. These results show no effects of exercise training on markers of hypercoagulability in our population with combined type 2 diabetes mellitus and coronary artery disease. The association between poor glycaemic control and tissue factor pathway inhibitor might indicate increased endothelial activation. More pronounced hypercoagulability and increased tissue factor pathway inhibitor were demonstrated in patients with albuminuria.


Intravenous tenoxicam to treat acute renal colic: comparison with buscopan compositum.
Forty-seven patients with acute renal colic were treated with either tenoxicam 20 mg i.v. or buscopan compositum (hyoscine butylbromide 20 mg and dipyrone 2.5 g) i.v. in a double blind study. Renal colic was diagnosed with use of a general urine examination, intravenous urogram, ultrasonography or voiding of calculus. The severity of symptoms were assessed by a verbal six point scale. Results demonstrated that 80% of patients treated with tenoxicam and 72.7% of patients treated with buscopan compositum showed significant improvement after 1 hour. Sixty-two percent of the patients who showed initial response to buscopan compositum had pain relapse during next 24 hours and required rescue treatment with pethidine 100 mg i.m. None of the patients treated with tenoxicam i.v. had pain relapse. No side effects were reported with use of tenoxicam. It is concluded that tenoxicam i.v. was more effective than antispasmodics and has rapid onset of analgesia and prolonged action in the treatment of acute renal colic.


Electric organ discharge from electric eel facilitates DNA transformation into teleost larvae in laboratory conditions.
Electric eels (Electrophorus sp.) are known for their ability to produce electric organ discharge (EOD) reaching voltages of up to 860 V. Given that gene transfer via intense electrical pulses is a well-established technique in genetic engineering, we hypothesized that electric eels could potentially function as a gene transfer mechanism in their aquatic environment. To investigate this hypothesis, we immersed zebrafish larvae in water containing DNA encoding the green fluorescent protein (GFP) and exposed them to electric eel's EOD. Some embryos exhibited a mosaic expression of green fluorescence, in contrast to the control group without electrical stimulation, which showed little distinct fluorescence. This suggests that electric eel EOD has the potential to function as an electroporator for the transfer of DNA into eukaryotic cells. While electric eel EOD is primarily associated with behaviors related to sensing, predation, and defense, it may incidentally serve as a possible mechanism for gene transfer in natural environment. This investigation represents the initial exploration of the uncharted impact of electric eel EOD, but it does not directly establish its significance within the natural environment. Further research is required to understand the ecological implications of this phenomenon.


Humoral and cellular immunogenicity to a second dose of COVID-19 vaccine BNT162b2 in people receiving methotrexate or targeted immunosuppression: a longitudinal cohort study.
COVID-19 vaccines have robust immunogenicity in the general population. However, data for individuals with immune-mediated inflammatory diseases who are taking immunosuppressants remains scarce. Our previously published cohort study showed that methotrexate, but not targeted biologics, impaired functional humoral immunity to a single dose of COVID-19 vaccine BNT162b2 (Pfizer-BioNTech), whereas cellular responses were similar. Here, we aimed to assess immune responses following the second dose. In this longitudinal cohort study, we recruited individuals with psoriasis who were receiving methotrexate or targeted biological monotherapy (ie, tumour necrosis factor [TNF] inhibitors, interleukin [IL]-17 inhibitors, or IL-23 inhibitors) from a specialist psoriasis centre serving London and South-East England. The healthy control cohort were volunteers without psoriasis, not receiving immunosuppression. Immunogenicity was evaluated immediately before, on day 28 after the first BNT162b2 vaccination and on day 14 after the second dose (administered according to an extended interval regimen). Here, we report immune responses following the second dose. The primary outcomes were humoral immunity to the SARS-CoV-2 spike glycoprotein, defined as titres of total spike-specific IgG and of neutralising antibody to wild-type, alpha (B.1.1.7), and delta (B.1.617.2) SARS-CoV-2 variants, and cellular immunity defined as spike-specific T-cell responses (including numbers of cells producing interferon-γ, IL-2, IL-21). Between Jan 14 and April 4, 2021, 121 individuals were recruited, and data were available for 82 participants after the second vaccination. The study population included patients with psoriasis receiving methotrexate (n=14), TNF inhibitors (n=19), IL-17 inhibitors (n=14), IL-23 inhibitors (n=20), and 15 healthy controls, who had received both vaccine doses. The median age of the study population was 44 years (IQR 33-52), with 43 (52%) males and 71 (87%) participants of White ethnicity. All participants had detectable spike-specific antibodies following the second dose, and all groups (methotrexate, targeted biologics, and healthy controls) demonstrated similar neutralising antibody titres against wild-type, alpha, and delta variants. By contrast, a lower proportion of participants on methotrexate (eight [62%] of 13, 95% CI 32-86) and targeted biologics (37 [74%] of 50, 60-85; p=0·38) had detectable T-cell responses following the second vaccine dose, compared with controls (14 [100%] of 14, 77-100; p=0·022). There was no difference in the magnitude of T-cell responses between patients receiving methotrexate (median cytokine-secreting cells per 106 cells 160 [IQR 10-625]), targeted biologics (169 [25-503], p=0·56), and controls (185 [133-328], p=0·41). Functional humoral immunity (ie, neutralising antibody responses) at 14 days following a second dose of BNT162b2 was not impaired by methotrexate or targeted biologics. A proportion of patients on immunosuppression did not have detectable T-cell responses following the second dose. The longevity of vaccine-elicited antibody responses is unknown in this population. NIHR Biomedical Research Centre at Guy's and St Thomas' NHS Foundation Trust and King's College London; The Psoriasis Association.


Electronic nose technology for detection of invasive pulmonary aspergillosis in prolonged chemotherapy-induced neutropenia: a proof-of-principle study.
Although the high mortality rate of pulmonary invasive aspergillosis (IA) in patients with prolonged chemotherapy-induced neutropenia (PCIN) can be reduced by timely diagnosis, a diagnostic test that reliably detects IA at an early stage is lacking. We hypothesized that an electronic nose (eNose) could fulfill this need. An eNose can discriminate various lung diseases through the analysis of exhaled volatile organic compounds (VOCs). An eNose is cheap and noninvasive and yields results within minutes. In a single-center prospective cohort study, we included patients who were treated with chemotherapy expected to result in PCIN. Based on standardized indications, a full diagnostic workup was performed to confirm invasive aspergillosis or to rule it out. Patients with no aspergillosis were considered controls, and patients with probable or proven aspergillosis were considered index cases. Exhaled breath was examined with a Cyranose 320 (Smith Detections, Pasadena, CA). The resulting data were analyzed using principal component reduction. The primary endpoint was cross-validated diagnostic accuracy, defined as the percentage of patients correctly classified using the leave-one-out method. Accuracy was validated by 100,000 random classifications. We included 46 subjects who underwent 16 diagnostic workups, resulting in 6 cases and 5 controls. The cross-validated accuracy of the eNose in diagnosing IA was 90.9% (P = 0.022; sensitivity, 100%; specificity, 83.3%). Receiver operating characteristic analysis showed an area under the curve of 0.93. These preliminary data indicate that PCIN patients with IA have a distinct exhaled VOC profile that can be detected with eNose technology. The diagnostic accuracy of the eNose for invasive aspergillosis warrants validation.


Transcriptomic Analyses and Potential Therapeutic Targets of Pancreatic Cancer With Concomitant Diabetes.
Type 2 diabetes mellitus (T2DM) known as non-insulin-dependent diabetes mellitus, which is increasingly acknowledged as being associated with an increased risk for a series of cancers. Pancreatic cancer is currently the fourth most common cause of cancer-related mortality, which has been proved to be worsened by internal diabetic condition. However, the underlying molecular mechanisms are less addressed. Furthermore, current knowledge revealed that therapeutic strategy by anti-diabetes for pancreatic cancer under diabetes condition have no satisfactory efficacy, and nor by chemotherapy in our study. To clarify these mysteries and widen our knowledge, both obesity-associated and non-obese-associated T2DM mouse models were generated by chemical induction with streptozotocin (STZ) and leptin receptor knockout (db/db) in mice. Then, the process of tumor progression was researched, and the gene expression profiling of pancreatic cancer in mice was performed using RNA-seq. Our results showed that pancreatic cancer malignancy was increased with notable proliferation and metastatic potential in two diabetic mice model. Totally, 136 and 64 significantly differentially expressed genes (DEGs) were identified in STZ and db/db mice by transcriptomic analysis. The results also suggested that different carcinogenesis-related genes and potential molecular mechanisms contribute to the malignancy of pancreatic cancer in obesity-associated and non-obesity-associated T2DM. In obesity-associated db/db mice, the GO subcategories associated with most of the genes with downregulated expression are involved in the immune response. However, in non-obesity-associated STZ mice, in addition to the immune response category, the enriched subcategories also included angiogenesis and the extracellular matrix. While, two genes respectively encoding MMP-2 and MMP-9 were simultaneously abnormal up-regulated in pancreatic cancer tissue from diabetic mice of both STZ and db/db, that could act as potential therapeutic targets for significantly suppressing the malignant progression. Furthermore, an optimizing therapeutic strategy was further proposed that combining MMP-2/9 inhibitor with gemcitabine significantly enhanced anti-tumor effects on pancreatic cancer under diabetic condition, providing a theoretical basis for clinical applications. Generally, this study provides a comprehensive insight into diabetes as a risk factor for pancreatic cancer and has the potential to guide the development of enhanced treatment strategies.


Cross-Talk Between Sporophyte and Gametophyte Generations Is Promoted by CHD3 Chromatin Remodelers in Arabidopsis thaliana.
Angiosperm reproduction requires the integrated development of multiple tissues with different genotypes. To achieve successful fertilization, the haploid female gametophytes and diploid ovary must coordinate their development, after which the male gametes must navigate through the maternal sporophytic tissues to reach the female gametes. After fertilization, seed development requires coordinated development of the maternal diploid integuments, the triploid endosperm, and the diploid zygote. Transcription and signaling factors contribute to communication between these tissues, and roles for epigenetic regulation have been described for some of these processes. Here we identify a broad role for CHD3 chromatin remodelers in Arabidopsis thaliana reproductive development. Plants lacking the CHD3 remodeler, PICKLE, exhibit various reproductive defects including abnormal development of the integuments, female gametophyte, and pollen tube, as well as delayed progression of ovule and embryo development. Genetic analyses demonstrate that these phenotypes result from loss of PICKLE in the maternal sporophyte. The paralogous gene PICKLE RELATED 2 is preferentially expressed in the endosperm and acts antagonistically with respect to PICKLE in the seed: loss of PICKLE RELATED 2 suppresses the large seed phenotype of pickle seeds. Surprisingly, the alteration of seed size in pickle plants is sufficient to determine the expression of embryonic traits in the seedling primary root. These findings establish an important role for CHD3 remodelers in plant reproduction and highlight how the epigenetic status of one tissue can impact the development of genetically distinct tissues.


A comprehensive framework for thermoeconomic analysis of desalination systems.
Thermoeconomic analysis, a combined application of thermodynamic and economic analyses, has emerged as an important tool to optimize the performance of desalination systems. Contrary to conventional economic analysis, it offers flexibility to investigate and improve the performance of each component in the system, individually. The current paper presents a comprehensive framework for conducting thermoeconomic analysis of desalination systems. In this regard, different energy calculation methods are discussed first. Then a detailed review of theoretical developments of thermoeconomic analyses is conducted to summarize the correlations/magnitude of important economic parameters. This is followed by a discussion on cost balance equations for important desalination components. Finally, a systematic thermoeconomic analysis model is developed for the mechanical vapor compression desalination system operating under different arrangements as an example. The monetary value of each stream calculated using appropriate fiscal parameters in the system is presented in the form of a cost flow diagram. The study can be used to conduct the thermoeconomic analysis of other commercial desalination systems.


Rhodium-catalyzed Sommelet-Hauser type rearrangement of α-diazoimines: synthesis of functionalized enamides.
An efficient rhodium catalyzed Sommelet-Hauser type rearrangement of sulfur ylides derived from α-thioesters and N-sulfonyl-1,2,3-triazoles has been successfully accomplished for the synthesis of various functionalized enamides. The developed reaction involves the unprecedented [2,3]-sigmatropic rearrangement of sulfur ylides with the imine motif. Importantly, the method works well with various substituted α-thioesters/-amides/-ketones and substituted N-sulfonyl-1,2,3-triazoles and allows the synthesis of diverse enamide derivatives in good to excellent yields. The reaction was also successfully extended to the one-pot synthesis of enamides from terminal alkynes.


Dacryoscintigraphy: an effective tool in the evaluation of postoperative epiphora.
Dacryoscintigraphy is a grossly underutilized nuclear medicine procedure for assessing the patency of nasolacrimal ducts. It is an alternate noninvasive modality requiring no added expertise when compared with dacryocystography, an invasive radiological counterpart. Dacryocystorhinostomy (DCR) is a surgical procedure in which a patent rhinostomy is made so that a low-pressure lacrimal bypass system is created, thus relieving epiphora, dacryocystitis or mucocele. DCR has been accepted as a highly successful procedure in the management of epiphora from nasolacrimal duct obstruction; however, adverse local effects can reproduce epiphora. This study highlights the utility and simplicity of dacryoscintigraphy in the postoperative setting of DCR, apart from showcasing its high sensitivity and accuracy in localizing the sites of lacrimal drainage obstruction. Fifty-five postoperative patients of DCR were screened. Ten volunteers constituted the control group. One drop (10 μl) of technetium pertechnetate (TcO4) was instilled simultaneously in the outer canthus of both eyes (50-100 μCi). Upright dynamic (8 min) and anterior static images of the eyes (up to 20 min) were acquired on a gamma camera fitted with a high-resolution collimator. Physiological interventions that were used in this study were eye blinking, saline intervention, and lacrimal sac massaging. Out of 40 symptomatic DCR patients, 22 had complete obstruction at the presac level and 18 patients had partial obstruction. Quantitative parameters like tear transit time and lacrimal sac half-life were also calculated. After massage of the lacrimal sac, progression of the tracer was observed in five patients. Premassage and postmassage tear column measurements showed a relative increase of 30.1%. Massage of eight clinically normal ducts showed a relative increase of 2.4% (P=0.06). Statistical analysis using the Fisher test shows very significant concordance between clinical symptoms and dacryoscintigraphy findings in postoperative DCR patients (P<0.0001). The modified McNemar's pair test did not show statistically significant differences between dacryoscintigraphy and the syringing irrigation test. Dacryoscintigraphy is a simple and easy-to-perform investigation providing high sensitivity in epiphora detection in patients after DCR. Interventions play a role in further enhancing the sensitivity of the technique. This study demonstrates the utility of dacryoscintigraphy in detecting subclinical and partial lacrimal duct obstruction with good patient compliance. It provides an easy and effective way of screening families, enabling an early diagnosis. During imaging, a high-resolution collimator is found to be an efficient substitute for the conventional pinhole collimator.


BAM8-22 peptide produces itch and nociceptive sensations in humans independent of histamine release.
Chronic itch accompanying many dermatological, neurological, and systemic diseases is unresponsive to antihistamines. Our knowledge of endogenous chemicals that evoke histamine-independent itch and their molecular targets is very limited. Recently it was demonstrated in behavioral and cellular experiments that bovine adrenal medulla 8-22 peptide (BAM8-22), a proteolytically cleaved product of proenkephalin A, is a potent activator of Mas-related G-protein-coupled receptors (Mrgprs), MrgprC11 and hMrgprX1, and induces scratching in mice in an Mrgpr-dependent manner. To study the sensory qualities that BAM8-22 evokes in humans, we tested the volar forearm of 15 healthy volunteers with heat-inactivated cowhage spicules previously soaked in the peptide. BAM8-22 produced itch in each subject, usually accompanied by sensations of pricking/stinging and burning. The sensations were occasionally accompanied by one or more mechanically evoked dysesthesias, namely alloknesis, hyperknesis, and/or hyperalgesia, but no wheal or neurogenic flare in the skin surrounding the application site. The inactive truncated peptide BAM8-18 produced weak or no sensations. Pretreatment of the tested skin with an antihistamine cream (doxepin) inhibited histamine-induced sensations, dysesthesias, and skin reactions but not the sensations and dysesthesias evoked by BAM8-22. We show that BAM8-22 produces itch and nociceptive sensations in humans in a histamine-independent manner. Thus, BAM8-22 may be an endogenous itch mediator that activates, in humans, MrgprX1, a novel target for potential anti-itch treatments.


Amyloid imaging in the differential diagnosis of dementia: review and potential clinical applications.
In the past decade, positron emission tomography (PET) with carbon-11-labeled Pittsburgh Compound B (PIB) has revolutionized the neuroimaging of aging and dementia by enabling in vivo detection of amyloid plaques, a core pathologic feature of Alzheimer's disease (AD). Studies suggest that PIB-PET is sensitive for AD pathology, can distinguish AD from non-AD dementia (for example, frontotemporal lobar degeneration), and can help determine whether mild cognitive impairment is due to AD. Although the short half-life of the carbon-11 radiolabel has thus far limited the use of PIB to research, a second generation of tracers labeled with fluorine-18 has made it possible for amyloid PET to enter the clinical era. In the present review, we summarize the literature on amyloid imaging in a range of neurodegenerative conditions. We focus on potential clinical applications of amyloid PET and its role in the differential diagnosis of dementia. We suggest that amyloid imaging will be particularly useful in the evaluation of mildly affected, clinically atypical or early age-at-onset patients, and illustrate this with case vignettes from our practice. We emphasize that amyloid imaging should supplement (not replace) a detailed clinical evaluation. We caution against screening asymptomatic individuals, and discuss the limited positive predictive value in older populations. Finally, we review limitations and unresolved questions related to this exciting new technique.


Delayed administration of interleukin-1 receptor antagonist reduces ischemic brain damage and inflammation in comorbid rats.
Many neuroprotective agents have been effective in experimental stroke, yet few have translated into clinical application. One reason for this may be failure to consider clinical comorbidities/risk factors in experimental models. We have shown that a naturally occurring interleukin-1 receptor antagonist (IL-1Ra) is protective against ischemic brain damage in healthy animals. However, protective effects of IL-1Ra have not been determined in comorbid animals. Thus, we tested whether IL-1Ra protects against brain injury induced by experimental ischemia in aged JCR-LA (corpulent) rats, which have clinically relevant risk factors. Male, aged, lean, and corpulent rats exposed to transient (90 minutes) occlusion of the middle cerebral artery (tMCAO) were administered two doses of IL-1Ra (25 mg/kg, subcutaneously) during reperfusion. Brain injury and neuroinflammatory changes were assessed 24 hours after tMCAO. Our results show that IL-1Ra administered at reperfusion significantly reduced infarct volume measured by magnetic resonance imaging (50%, primary outcome) and blood-brain barrier disruption in these comorbid animals. Interleukin-1Ra also reduced microglial activation, neutrophil infiltration, and cytokines levels in the brain. These data are the first to indicate that IL-1Ra protects against ischemic brain injury when administered via a clinically relevant route and time window in animals with multiple risk factors for stroke.


Developmental regulation of lifelong dental experiences and beliefs in Guangzhou and Hong Kong.
The aim of this study was to explain through the life-course and life-span perspectives of developmental regulation theory the controls on dental experiences and beliefs throughout the lives of older people in Guangzhou and Hong Kong. Dental diseases and disabilities among older people are serious public health concerns in China. A facilitator conducted eight focus groups, three in Hong Kong and five in Guangzhou, involving a total of 51 participants. She encouraged discussions about lifetime events to explain dental experiences and beliefs. Transcripts were coded and analysed using a constant comparative approach to identify themes that explained the regulators of dental experiences throughout the participants' lives. Participants explained the influence of culture and history through critical events, and how external and internal factors regulated their current oral health status and beliefs. They emphasised the role of Traditional Chinese Medicine and family, and the stress of social upheaval compounded by a scarcity of dental services. They revealed also how current choice of dental services and health promotional programs, helped by personal food choice, self-reliance, and scepticism, helped them to adjust and cope with dental diseases and disabilities and the commercialisation of dental services. Dental experiences and beliefs of older people living in Guangzhou and Hong Kong were regulated strongly during personal development by culture and history during critical events, and by various controlling factors, such as health promotion and choice of services supplemented by food choice, nutritional balance, self-reliance, scepticism and social adjustments.


Dissection of floral induction pathways using global expression analysis.
Flowering of the reference plant Arabidopsis thaliana is controlled by several signaling pathways, which converge on a small set of genes that function as pathway integrators. We have analyzed the genomic response to one type of floral inductive signal, photoperiod, to dissect the function of several genes transducing this stimulus, including CONSTANS, thought to be the major output of the photoperiod pathway. Comparing the effects of CONSTANS with those of FLOWERING LOCUS T, which integrates inputs from CONSTANS and other floral inductive pathways, we find that expression profiles of shoot apices from plants with mutations in either gene are very similar. In contrast, a mutation in LEAFY, which also acts downstream of CONSTANS, has much more limited effects. Another pathway integrator, SUPPRESSOR OF OVEREXPRESSION OF CO 1, is responsive to acute induction by photoperiod even in the presence of the floral repressor encoded by FLOWERING LOCUS C. We have discovered a large group of potential floral repressors that are down-regulated upon photoperiodic induction. These include two AP2 domain-encoding genes that can repress flowering. The two paralogous genes, SCHLAFMUTZE and SCHNARCHZAPFEN, share a signature with partial complementarity to the miR172 microRNA, whose precursor we show to be induced upon flowering. These and related findings on SPL genes suggest that microRNAs play an important role in the regulation of flowering.


Not all sharks are "swimming noses": variation in olfactory bulb size in cartilaginous fishes.
Olfaction is a universal modality by which all animals sample chemical stimuli from their environment. In cartilaginous fishes, olfaction is critical for various survival tasks including localizing prey, avoiding predators, and chemosensory communication with conspecifics. Little is known, however, about interspecific variation in olfactory capability in these fishes, or whether the relative importance of olfaction in relation to other sensory systems varies with regard to ecological factors, such as habitat and lifestyle. In this study, we have addressed these questions by directly examining interspecific variation in the size of the olfactory bulbs (OB), the region of the brain that receives the primary sensory projections from the olfactory nerve, in 58 species of cartilaginous fishes. Relative OB size was compared among species occupying different ecological niches. Our results show that the OBs maintain a substantial level of allometric independence from the rest of the brain across cartilaginous fishes and that OB size is highly variable among species. These findings are supported by phylogenetic generalized least-squares models, which show that this variability is correlated with ecological niche, particularly habitat. The relatively largest OBs were found in pelagic-coastal/oceanic sharks, especially migratory species such as Carcharodon carcharias and Galeocerdo cuvier. Deep-sea species also possess large OBs, suggesting a greater reliance on olfaction in habitats where vision may be compromised. In contrast, the smallest OBs were found in the majority of reef-associated species, including sharks from the families Carcharhinidae and Hemiscyllidae and dasyatid batoids. These results suggest that there is great variability in the degree to which these fishes rely on olfactory cues. The OBs have been widely used as a neuroanatomical proxy for olfactory capability in vertebrates, and we speculate that differences in olfactory capabilities may be the result of functional rather than phylogenetic adaptations.


Structural analysis of phospholipase A2 from functional perspective. 1. Functionally relevant solution structure and roles of the hydrogen-bonding network.
Bovine pancreatic phospholipase A2 (PLA2), a small (13.8 kDa) Ca2+-dependent lipolytic enzyme, is rich in functional and structural character. In an effort to examine its detailed structure-function relationship, we determined its solution structure by multidimensional nuclear magnetic resonance (NMR) spectroscopy at a functionally relevant pH. An ensemble of 20 structures generated has an average root-mean-square deviation (RMSD) of 0.62 +/- 0.08 A for backbone (N, Calpha, C) atoms and 0.98 +/- 0.09 A for all heavy atoms. The overall structure shows several notable differences from the crystal structure: the first three residues at the N-terminus, the calcium-binding loop (Y25-T36), and the surface loop (V63-N72) appear to be flexible; the alpha-helical conformation of helix B (E17-F22) is absent; helix D appears to be shorter (D59-V63 instead of D59-D66); and the hydrogen-bonding network is less defined. These differences were analyzed in relation to the function of PLA2. We then further examined the H-bonding network, because its functional role or even its existence in solution has been in dispute recently. Our results show that part of the H-bonding network (the portion away from N-terminus) clearly exists in solution, as evidenced by direct observation (at 11.1 ppm) of a strong H-bond between Y73 and D99 and an implicated interaction between D99 and H48. Analyses of a series of mutants indicated that the existence of the Y73.D99 H-bond correlates directly with the conformational stability of the mutant. Loss of this H-bond results in a loss of 2-3 kcal/mol in the conformational stability of PLA2. The unequivocal identification and demonstration of the structural importance of a specific hydrogen bond, and the magnitude of its contribution to conformational stability, are uncommon to the best of our knowledge. Our results also suggest that, while the D99.H48 catalytic diad is the key catalytic machinery of PLA2, it also helps to maintain conformational integrity.


Relevance of Geriatric Assessment in Older Patients With Colorectal Cancer.
This study aims to evaluate the relevance of geriatric assessment (GA) in older patients with colorectal cancer (CRC) and to study functional status (FS) and chemotherapy-related toxicity during treatment. Patients with CRC aged ≥ 70 years were evaluated at baseline using a GA. Results were communicated to the treating physician. At 2 to 3 months follow-up, FS was reassessed, and chemotherapy-related toxicity was recorded. A total of 193 patients, with a median age of 77 years, were included. GA was abnormal in 75% and revealed unknown problems in 40%. Treatment was altered in 37% based on clinical assessment. GA led to geriatric interventions in 9 patients (5%) and additionally influenced treatment in 1 patient. At follow-up (n = 164), functional decline was observed in 29 patients (18%) for activities of daily living (ADL) and in 60 patients (37%) for instrumental activities of daily living (IADL). Baseline IADL, depression, fatigue, and cognition were predictors for ADL decline, whereas no predictors for IADL decline could be identified. In the 109 patients receiving chemotherapy, stage and baseline fatigue were predictive for grade 3/4 hematologic toxicity, and baseline ADL, fatigue, and nutrition were predictive for grade 3/4 nonhematologic toxicity. Although GA identified previously unknown problems in more than one-third of older CRC patients, the impact on interventions or treatment decisions was limited. Baseline GA parameters may predict functional decline and chemotherapy-related toxicity. Education of physicians treating older patients with CRC is an essential step in the implementation of GA and subsequent interventions.


Protection against focal ischemic injury to the brain by trans-sodium crocetinate. Laboratory investigation.
Ischemic injury is a potential complication in a variety of surgical procedures and is a particular impediment to the success of surgeries involving highly vulnerable neural tissue. One approach to limiting this form of injury is to enhance metabolic supply to the affected tissue. Trans-sodium crocetinate (TSC) is a carotenoid compound that has been shown to increase tissue oxygenation by facilitating the diffusivity of small molecules, such as oxygen and glucose. The present study examined the ability of TSC to modify oxygenation in ischemic neural tissue and tested the potential neuroprotective effects of TSC in permanent and temporary models of focal cerebral ischemia. Adult male rats (330–370 g) were subjected to either permanent or temporary focal ischemia by simultaneous occlusion of both common carotid arteries and the left middle cerebral artery (3-vessel occlusion [3-VO]). Using the permanent ischemia paradigm, TSC was administered intravenously beginning 10 minutes after the onset of ischemia at 1 of 8 dosages, ranging from 0.023 to 4.580 mg/kg. Cerebral infarct volume was measured 24 hours after the onset of ischemia. The effect of TSC on infarct volume was also tested after temporary (2-hour) ischemia using a dosage of 0.092 mg/kg. In other animals undergoing temporary ischemia, tissue oxygenation was monitored in the ischemic penumbra using a Licox probe. Administration of TSC reduced infarct volume in a dose-dependent manner in the permanent ischemia model, achieving statistical significance at dosages ranging from 0.046 to 0.229 mg/kg. The most effective dosage of TSC in the permanent ischemia experiment (0.092 mg/kg) was further tested using a temporary (2-hour) ischemia paradigm. Infarct volume was reduced significantly by TSC in this ischemia-reperfusion model as well. Recordings of oxygen levels in the ischemic penumbra of the temporary ischemia model showed that TSC increased tissue oxygenation during vascular occlusion, but reduced the oxygen overshoot (hyperoxygenation) that occurs upon reperfusion. The novel carotenoid compound TSC exerts a neuroprotective influence against permanent and temporary ischemic injury when administered soon after the onset of ischemia. The protective mechanism of TSC remains to be confirmed; however, the permissive effect of TSC on the diffusivity of small molecules is a plausible mechanism based on the observed increase in tissue oxygenation in the ischemic penumbra. This represents a form of protection based on "metabolic reflow" that can occur under conditions of partial vascular perfusion. It is particularly noteworthy that TSC could conceivably limit the progression of a wide variety of cellular injury mechanisms by blunting the ischemic challenge to the brain.


A Case of Segmental Aplasia of the Uterus, Cervix, and Cranial Vagina in a Cat.
This case documents a rare set of congenital anomalies that resulted in an atypical cystic lesion in the cranial vagina of a queen. A discrete cystic lesion was identified in an 8 year old intact female domestic shorthair cat presenting for routine ovariohysterectomy. Morphological, radiographic, and histopathological findings were consistent with segmental aplasia of the uterus, cervix, and vagina resulting in a blind dilation of the cranial vagina. Segmental vaginal aplasia in combination with the failed canalization of the cervix resulted in a blind portion of the cranial vagina, in which normal secretions collected and became inspissated. This formed a discrete cystic structure. This case represents a novel combination and clinical presentation of segmental aplasia in the cat, involving both the uterus and cranial vagina. Ovariohysterectomy was performed for sterilization and a partial vaginectomy was performed to remove the cystic lesion in its entirety.


Stakeholder consultations on community-based rehabilitation guidelines in Ghana and Uganda.
The focus of this paper is the new broadened conceptualisation of community-based rehabilitation (CBR), which promotes the empowerment and inclusion of people with disabilities (PWDs) in diverse ways within their communities. New guidelines for CBR were launched in October 2010 by WHO/ILO/UNESCO/IDDC, and this paper describes part of the process by which these were produced using participatory approaches involving International Non-Government Organisations (INGOs) and local partners. The paper reviews the evolution of CBR and describes how grassroots consultation by INGOs working with key stakeholders in the disability arena can influence policy on disability issues, and reciprocally how policy change can inform organisations' practice and research activities. This ongoing bidirectional influence is illustrated with data from the participatory consultation process about the new CBR guidelines carried out by Sightsavers in Uganda and Ghana. To consult with key stakeholders in the disability arena in Uganda and Ghana, in order to gain their opinions and suggestions for improvements to the then draft CBR guidelines, as part of a wider global participatory process of consultation on the document. The INGO Sightsavers gathered qualitative data through focus group discussions and questionnaires in both countries. The participants' critiques of the draft guidelines carried out in multiagency participatory processes were analysed thematically and fed back to the CBR guidelines editorial team. The paper concludes that stakeholders in diverse communities can actively contribute to shaping policy and practice through participatory consultations. Local and national government and non-government organisations and other key informants can inform the development of national and international guidelines and policies. This participatory approach can be successfully facilitated by INGOs. In turn, these processes have prompted organisations to adapt their own policies and programmes in order to be more responsive to the local needs and concerns of PWDs.


[Apheresis in inflammatory bowel disease. A valid option?].
Leukocyte apheresis works by purifying distinct populations of active leukocytes and by modifying inflammatory mediators. Based on their theoretical immunomodulatory effect, these techniques have begun to be used in inflammatory bowel disease. There are two types of devices: granulocytapheresis, which performs selective granulocyte and monocyte absorption, and leukocytapheresis, which carries out non-selective filtration. Conventional treatment regimens consist of between 5 and 10 sessions, normally one session per week, to induce remission. Although the indications for apheresis have not been clearly defined, this technique can be considered a valid option in selected patients with ulcerative colitis, mainly those with corticosteroid-dependent or corticosteroid-refractory disease when other, better-established immunosuppressive or biological treatments have failed. In addition to avoiding the use of corticosteroids, due to its excellent safety profile, apheresis is an attractive option to avoid the risks of immunosuppressive and biological treatments and for use in the pediatric population. The present reviews analyzes the data on the safety and efficacy of apheresis in those patients with inflammatory bowel diseases who could benefit from this technique.


Pseudomonas Prosthetic Joint Infections: A Review of 102 Episodes.
Background: The outcome of patients with Pseudomonas prosthetic joint infection (PS PJI) has not been well studied. The aim of this retrospective cohort study was to assess the outcome of patients with Pseudomonas PJI and to review risk factors associated with failure of therapy. Methods: Between 1/1969 and 12/2012, 102 episodes of PS PJI in 91 patients were identified. Results: The mean age at the time of diagnosis was 67.4 years; forty three percent had knee involvement. Over 40 percent had either diabetes mellitus or a history of gastrointestinal or genitourinary surgery. Nearly half (48 out of 102 episodes) received aminoglycoside monotherapy, while 25% received an anti-pseudomonal cephalosporin. The 2-year cumulative survival free from failure was 69% (95% CI, 56%-82%). Patients treated with resection arthroplasty, two-stage exchange, and debridement with implant retention had a 2-year cumulative survival free from failure of 80% (95% CI, 66%-95%), 83% (95% CI, 60%-100%), and 26% (95% CI, 23%-29%) respectively (P=0.0001). Conclusions: PS PJI's are associated with a high failure rate. Patients treated with debridement and implant retention had a worse outcome.


Increased Myocardial Oxygen Consumption Precedes Contractile Dysfunction in Hypertrophic Cardiomyopathy Caused by Pathogenic TNNT2 Gene Variants.
Background Hypertrophic cardiomyopathy is caused by pathogenic sarcomere gene variants. Individuals with a thin-filament variant present with milder hypertrophy than carriers of thick-filament variants, although prognosis is poorer. Herein, we defined if decreased energetic status of the heart is an early pathomechanism in TNNT2 (troponin T gene) variant carriers. Methods and Results Fourteen individuals with TNNT2 variants (genotype positive), without left ventricular hypertrophy (G+/LVH-; n=6) and with LVH (G+/LVH+; n=8) and 14 healthy controls were included. All participants underwent cardiac magnetic resonance and [11C]-acetate positron emission tomography imaging to assess LV myocardial oxygen consumption, contractile parameters and myocardial external efficiency. Cardiac efficiency was significantly reduced compared with controls in G+/LVH- and G+/LVH+. Lower myocardial external efficiency in G+/LVH- is explained by higher global and regional oxygen consumption compared with controls without changes in contractile parameters. Reduced myocardial external efficiency in G+/LVH+ is explained by the increase in LV mass and higher oxygen consumption. Septal oxygen consumption was significantly lower in G+/LVH+ compared with G+/LVH-. Although LV ejection fraction was higher in G+/LVH+, both systolic and diastolic strain parameters were lower compared with controls, which was most evident in the hypertrophied septal wall. Conclusions Using cardiac magnetic resonance and [11C]-acetate positron emission tomography imaging, we show that G+/LVH- have an initial increase in oxygen consumption preceding contractile dysfunction and cardiac hypertrophy, followed by a decline in oxygen consumption in G+/LVH+. This suggests that high oxygen consumption and reduced myocardial external efficiency characterize the early gene variant-mediated disease mechanisms that may be used for early diagnosis and development of preventive treatments.


Event-related oscillations (ERO) and event-related potentials (ERP) in emotional face recognition.
The study aims to explore the significance of event-related potentials (ERPs) and event-related brain oscillations (EROs) (delta, theta, and alpha power) in response to emotional face during 180-250 poststimulus time interval. Twenty-one adults looked at emotional (sad, happy, fearful) or neutral faces. The results demonstrated that the emotional face elicited a negative peak at approximately 230 ms (N230). Moreover EEG measures showed that motivational significance of face (stimulus type) can modulate the amplitude of EEG, especially for theta and delta. Regression analysis showed that theta oscillations mainly effect as oscillation activity at the N2 latency. Thus, this frequency band variation could represent a complex set of cognitive processes, whereby selective attention becomes focused on an emotional relevant stimulus.


Carotid intima-media thickness and serum proinflammatory cytokine levels in rosacea patients without cardiovascular risk factors.
There is a growing body of evidence linking rosacea to various systemic disorders, even though data regarding the association between rosacea and cardiovascular diseases are presently controversial. We sought to investigate the potential association of rosacea with subclinical atherosclerosis and serum proinflammatory/proatherogenic markers. This study included 44 patients with rosacea and 44 age-matched and sex-matched healthy control subjects. Patients with traditional cardiovascular risk factors or a history of cardiovascular events were excluded. Demographic, clinical, and laboratory data, including serum interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and high-sensitivity C-reactive protein (hs-CRP) levels were assessed. Carotid intima-media thickness (CIMT) and carotid plaques were measured by carotid ultrasonography. Serum IL-1β (P < .001), IL-6 (P < .001), TNF-α (P < .001), and hs-CRP (P < .001) levels were significantly higher in the patient group compared with the control group. Mean CIMT values did not differ significantly between the patient group and control group (P > .05). Patients with moderate to severe rosacea had a significantly greater CIMT than those with mild rosacea (P = .047). Rosacea patients with eye involvement had a significantly greater CIMT than those without eye involvement (P = .008). There was no significant correlation between CIMT values and inflammation parameters. As conclusion, in the absence of other traditional cardiovascular risk factors, rosacea does not seem to affect mean CIMT value. However, specific subgroups such as patients with moderate to severe disease or with eye involvement are associated with increased subclinical atherosclerosis and may require additional attention for cardiovascular disease prevention.


Electron microscopy of the early Caenorhabditis elegans embryo.
The early Caenorhabditis elegans embryo is currently a popular model system to study centrosome assembly, kinetochore organization, spindle formation, and cellular polarization. Here, we present and review methods for routine electron microscopy and 3D analysis of the early C. elegans embryo. The first method uses laser-induced chemical fixation to preserve the fine structure of isolated embryos. This approach takes advantage of time-resolved fixation to arrest development at specific stages. The second method uses high-pressure freezing of whole worms followed by freeze-substitution (HPF-FS) for ultrastructural analysis. This technique allows staging of developing early embryos within the worm uterus, and has the advantage of superior sample preservation required for high-resolution 3D reconstruction. The third method uses a correlative approach to stage isolated, single embryos by light microscopy followed by HPF-FS and electron tomography. This procedure combines the advantages of time-resolved fixation and superior ultrastructural preservation by high-pressure freezing and allows a higher throughput electron microscopic analysis. The advantages and disadvantages of these methods for different applications are discussed.


Fine needle aspiration of chromophobe renal cell carcinoma.
To describe the cytomorphologic findings of chromophobe renal cell carcinoma (CRCC) in order to preoperatively distinguish this rare neoplasm from other primary or secondary tumors arising from the kidney or presenting as retroperitoneal masses. Clinical data, fine needle aspiration (FNA) and follow-up surgical specimens from 4 patients with CRCC (3 primaries and 1 metastatic to the liver) were reviewed. Electron microscopy was available for 2 histologic specimens. Two tumors (1 primary and 1 metastatic case) were readily identified as CRCC on FNA. The 2 remaining cases were diagnosed as renal cell carcinoma (RCC) consistent with CRCC. All tumors showed aspirates with moderate to high cellularity, with the cells arranged in small clusters and single cells. Neoplastic cells had abundant heterogeneous cytoplasm, a thickened cell membrane, nuclear hyperchromasia, nuclear outline irregularity, significant nuclear size variation, intranuclear inclusions and frequent binucleation. Histology of the 4 renal tumors was characteristic of CRCC, with positivity for Hale's colloidal iron in all cases. Ultrastructurally, characteristic cytoplasmic microvesicles were observed in the 2 cases that we studied. In the adequate clinicoradiologic setting, CRCC has distinctive cytologic features that may allow an accurate preoperative FNA diagnosis.


Impact of flow cytometry in liver cytopathology.
Fine needle aspiration (FNA) is commonly used as a diagnostic tool for the evaluation of lymphoproliferative diseases. Cytomorphology alone is often insufficient for the diagnosis and subclassification of lymphoma; therefore, flow cytometry (FC) plays an important role in the characterization of lymphoproliferative disorders. This study reviews our experience with FC on liver FNA at the Johns Hopkins Hospital. 2,424 liver FNAs performed over a 21-year period were reviewed for clinical FC data (n=74) or a subsequent diagnosis of lymphoma in the liver without FC data (n=40). In our study, 114 cases (4.7%) were included out of the 2,424 liver FNAs performed during the study period. Lymphoma was diagnosed 79 times. Cytomorphology alone was diagnostic of lymphoma in 45 cases, and in 33 cases both the cytomorphology and the FC were consistent with a diagnosis of lymphoma. Neither FC nor cytomorphology were diagnostic of lymphoma on 1 specimen. In 39 cases, FC had negative results on a lesion suspicious for lymphoma based on cytomorphology. In several nonlymphoma cases, FC provided information that allowed further subclassification of the neoplasm. FC is a useful adjuvant diagnostic test for liver FNAs performed on patients with lymphoproliferative disorders.


Impaired updating ability in drivers with Parkinson's disease.
Driving activity requires major involvement of executive functions. The main objective of our study was to determine whether mental flexibility and the updating of information in working memory are affected in drivers with mild to moderate Parkinson's disease (PD). The study included 25 patients, aged 58-76, with mild to moderate PD and 25 healthy controls matched for age, sex and education, with an average mileage of over 3000 km/year. Neuropsychological tests were conducted to assess global cognitive abilities, to evaluate updating (via the n-back task), flexibility (via the plus-minus task) and information-processing speed (via the Stroop test). Three different scenarios were developed on a driving simulator. Participants were asked to recall road signs (updating task), indicate the shape or colour of road signs according to road side (flexibility task) and to brake at the same time as the car ahead (information-processing speed task) while driving. An updating impairment was found in PD patients in the n-back and simulator tasks; patients recalled significantly fewer road signs. No notable differences were observed between groups in the plus-minus task or in the simulator task evaluating flexibility. There was no significant difference between patients and controls in information-processing speed tasks. Regression analysis showed that the Trail-Making test (B-A) accounted for 40.7% of the variation in PD drivers' simulator task updating score. The updating function is clearly impaired in drivers with mild to moderate PD, while mental flexibility remains unaffected. This study demonstrates the interest of using the Trail Making Test and simulator tasks to assess PD drivers.


Cerebellar medulloblastoma presenting with skeletal metastasis.
Medulloblastomas are highly malignant brain tumours, but only rarely produce skeletal metastases. No case of medulloblastoma has been documented to have produced skeletal metastases prior to craniotomy or shunt surgery. A 21-year-old male presented with pain in the hip and lower back with difficulty in walking of 3 months' duration. Signs of cerebellar dysfunction were present hence a diagnosis of cerebellar neoplasm or skeletal tuberculosis with cerebellar abscess formation was considered. MRI of brain revealed a lesion in the cerebellum suggestive of medulloblastoma. Bone scan revealed multiple sites of skeletal metastases excluding the lumbar vertebrae. MRI of lumbar spine and hip revealed metastases to all lumbar vertebrae and both hips. Computed tomography-guided biopsy was obtained from the L3 vertebra, which revealed metastatic deposits from medulloblastoma. Cerebrospinal fluid cytology showed the presence of medulloblastoma cells. A final diagnosis of cerebellar medulloblastoma with skeletal metastases was made. He underwent craniotomy and histopathology confirmed medulloblastoma.


"Cancer's a demon": a qualitative study of fear and multilevel factors contributing to cancer treatment delays.
Delays initiating cancer therapy are increasingly common, impact outcomes, and have implications for health equity. However, it remains unclear (1) whether patients' beliefs regarding acceptable diagnostic to treatment intervals align with current guidelines, and (2) to what degree psychological factors contribute to longer intervals. We conducted a qualitative study with patients and cancer care team members ("providers"). We interviewed patients with several common solid tumors as well as providers. Interviews were analyzed using an interpretive approach, guided by modified grounded theory. Twenty-two patients and 12 providers participated. Half of patients had breast cancer; 27% waited >60 days between diagnosis and treatment. Several themes emerged. (1) Patients felt treatment should begin immediately following diagnosis, while providers' opinion on the goal timeframe to start treatment varied. (2) Patients experienced psychological distress while waiting for treatment. (3) Participants identified logistical, social, and psychological sources of delay. Fear related to multiple aspects of cancer care was common. Emotion-driven barriers could manifest as not taking steps to move ahead, or as actions that delayed care. (4) Besides addressing logistical challenges, patients believed that education and anticipatory guidance, from their care team and from peers, may help overcome psychological barriers to treatment and facilitate the start of therapy. Patients feel an urgency to start cancer therapy, desiring time frames shorter than those included in guidelines. Psychological distress is frequently both a contributor to, and a consequence of, treatment delays. Addressing multilevel barriers, including psychological ones, may facilitate timely treatment and reduce distress.


Inulin Supplementation Does Not Reduce Plasma Trimethylamine N-Oxide Concentrations in Individuals at Risk for Type 2 Diabetes.
Trimethylamine N-oxide (TMAO) is associated with type 2 diabetes (T2DM) and increased risk of adverse cardiovascular events. Prebiotic supplementation has been purported to reduce TMAO production, but whether prebiotics reduce fasting or postprandial TMAO levels is unclear. Sedentary, overweight/obese adults at risk for T2DM (n = 18) were randomized to consume a standardized diet (55% carbohydrate, 30% fat) with 10 g/day of either an inulin supplement or maltodextrin placebo for 6 weeks. Blood samples were obtained in the fasting state and hourly during a 4-h high-fat challenge meal (820 kcal; 25% carbohydrate, 63% fat; 317.4 mg choline, 62.5 mg betaine, 8.1 mg l-carnitine) before and after the diet. Plasma TMAO and trimethylamine (TMA) moieties (choline, l-carnitine, betaine, and γ-butyrobetaine) were measured using isocratic ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). There were no differences in fasting or postprandial TMAO or TMA moieties between the inulin and placebo groups at baseline (all p > 0.05). There were no significant changes in fasting or postprandial plasma TMAO or TMA moiety concentrations following inulin or placebo. These findings suggest that inulin supplementation for 6 weeks did not reduce fasting or postprandial TMAO in individuals at risk for T2DM. Future studies are needed to identify efficacious interventions that reduce plasma TMAO concentrations.


Plasticity occurs in a specific phenotype of neurons in the nucleus tractus solitarius of dystrophin gene-mutated rats.
Duchenne muscular dystrophy (DMD) is a severe progressive neuromuscular disorder that causes cardiac and respiratory failure. Patients with DMD have tachycardia and autonomic nervous dysfunction at a young age, which can potentially worsen cardiorespiratory function. Therefore, we hypothesised that plasticity occurs in neurons of the cardiorespiratory brainstem nucleus (nucleus tractus solitarius [NTS]) due to DMD, thus affecting neuronal regulation because afferent information from cardiorespiratory organs changes with disease progression. Patch-clamp experiments were performed on second-order NTS neurons from Dmd-mutated (Dm) rats that showed no functional dystrophin protein expression, as confirmed by immunohistochemistry. NTS neurons are classified into two electrophysiological phenotypes: one showing a delayed onset of spiking from hyperpolarised membrane potentials, namely, delayed-onset spiking (DS)-type neurons, and the other showing a rapid onset, namely, rapid-onset spiking-type neurons. Neuroplasticity mainly occurs in DS-type neurons in Dm rats and is characterised by blunted neuronal excitability accompanied by reduced outward currents and a facilitatory effect on synaptic transmission, that is, an increased frequency of spontaneous and miniature excitatory postsynaptic currents (EPSCs) without changes in the amplitude and an increased amplitude of tractus solitarius-evoked EPSCs without changes in the paired-pulse ratio. Although we cannot rule out the possibility that the neuroplastic changes observed in Dm rats were caused by dystrophin deficiency in the neurons themselves, the plasticity could be caused by cardiorespiratory deterioration and/or adaptation in DMD patients.


An improved design for a stable and reproducible phantom material for use in near-infrared spectroscopy and imaging.
In this note, we describe an improved phantom material for use in near-infrared spectroscopy and imaging. The material consists of a clear epoxy resin with absorbing dyes and amorphous silica spheres as scattering particles. It is possible to calculate the scattering coefficient and angular scattering distribution of the material from Mie theory, using the known size and refractive index of the silica spheres together with the measured refractive index of the resin (approximately 1.56). We show a good agreement between prediction and experimental measurements. The scattering properties of the material closely match those of tissue in the near-infrared wavelength region, having an anisotropy factor, g, of approximately 0.93. The absorption coefficient of the epoxy is low (approximately 0.001 mm-1), and addition of the dyes produces an absorption coefficient that covers the same range as that of tissue.


Efficient and color-tunable Mn-doped ZnSe nanocrystal emitters: control of optical performance via greener synthetic chemistry.
Formation of Mn-doped ZnSe quantum dots (Mn:ZnSe d-dots) using nucleation-doping strategy was studied systematically and optimized through greener approaches. The resulting d-dots were with high ( approximately 50%) photoluminescence (PL) quantum yield (QY), which was achieved by the controlled formation of small-sized MnSe nanoclusters as the core and a diffused interface between the nanocluster core and the ZnSe overcoating layers. Synthesis of the d-dots under high temperatures (240-300 degrees C) was achieved by varying the structure of the metal carboxylate precursors, concentration of the inhibitors, free fatty acid, and concentration of the activation reagents, fatty amines. Highly emissive d-dots synthesized under desired conditions were found to be extremely stable upon thermal treatment up to the boiling point of the solvent (about 300 degrees C), which was quantitatively studied using in situ measurements. The PL peak of the d-dots was controllably tuned in a surprisingly large optical window, from 565 to 610 nm. These highly emissive and stable d-dots possess characteristics of practical emissive materials, especially for applications requiring high power, high concentration of emitters, and under tough conditions.


Silicon isotopes in Arctic and sub-Arctic glacial meltwaters: the role of subglacial weathering in the silicon cycle.
Glacial environments play an important role in high-latitude marine nutrient cycling, potentially contributing significant fluxes of silicon (Si) to the polar oceans, either as dissolved silicon (DSi) or as dissolvable amorphous silica (ASi). Silicon is a key nutrient in promoting marine primary productivity, contributing to atmospheric CO2 removal. We present the current understanding of Si cycling in glacial systems, focusing on the Si isotope (δ30Si) composition of glacial meltwaters. We combine existing glacial δ30Si data with new measurements from 20 sub-Arctic glaciers, showing that glacial meltwaters consistently export isotopically light DSi compared with non-glacial rivers (+0.16‰ versus +1.38‰). Glacial δ30SiASi composition ranges from -0.05‰ to -0.86‰ but exhibits low seasonal variability. Silicon fluxes and δ30Si composition from glacial systems are not commonly included in global Si budgets and isotopic mass balance calculations at present. We discuss outstanding questions, including the formation mechanism of ASi and the export of glacial nutrients from fjords. Finally, we provide a contextual framework for the recent advances in our understanding of subglacial Si cycling and highlight critical research avenues for assessing potential future changes in these environments.


Genetic method to identify regulons controlled by nonessential elements: isolation of a gene dependent on alternate transcription factor sigma B of Bacillus subtilis.
We describe a general, in vivo method for identifying Bacillus subtilis genes controlled by specific, nonessential regulatory factors. We establish the use of this approach by identifying, isolating, and characterizing a gene dependent on sigma B, an alternate transcription factor which is found early in stationary phase but which is not essential for sporulation. The method relies on two features: (i) a plate transformation technique to introduce a null mutation into the regulatory gene of interest and (ii) random transcriptional fusions to a reporter gene to monitor gene expression in the presence and absence of a functional regulatory product. We applied this genetic approach to isolate genes comprising the sigma B regulon. We screened a random Tn917lacZ library for fusions that required an intact sigma B structural gene (sigB) for greatest expression, converting the library strains from wild-type sigB+ to sigB delta::cat directly on plates selective for chloramphenicol resistance. We isolated one such fusion, csbA::Tn917lacZ (csb for controlled by sigma B), which mapped between hisA and degSU on the B. subtilis chromosome. We cloned the region surrounding the insertion, identified the csbA reading frame containing the transposon, and found that this frame encoded a predicted 76-residue product which was extremely hydrophobic and highly basic. Primer extension and promoter activity experiments identified a sigma B-dependent promoter 83 bp upstream of the csbA coding sequence. A weaker, tandem, sigma A-like promoter was likewise identified 28 bp upstream of csbA. The csbA fusion was maximally expressed during early stationary phase in cells grown in Luria broth containing 5% glucose and 0.2% glutamine. This timing of expression and medium dependence were very similar to those for ctc, the only other recognized gene dependent on sigma B.


The Great Island subgroup of tick-borne orbiviruses represents a single gene pool.
The geographical distribution of members of the Great Island (GI) subgroup in the Kemerovo serogroup of orbiviruses extends from the Arctic to the Sub-antarctic. To examine the gene pool size of this group, five topotypes whose origins ranged from Iceland in the northern hemisphere to Macquarie Island in the Southern Ocean were tested for their ability to reassort in vitro. All the isolates were distinguishable by plaque reduction neutralization tests, and their genome profile in polyacrylamide gels. They showed high frequency reassortment following dual infection of cell cultures with temperature-sensitive (ts) mutants and/or wild-type virus. Analysis of the dsRNA profile of the reassortants by PAGE confirmed the observation from reassortment assays that the Great Island subgroup constitutes a single gene pool. A seventh reassortment group was identified, distinct from the six groups previously described. The ts lesions for reassortment groups I, V and VII were considered to be in genome segments 9, 3 and 2, respectively. Segment 6 of GI virus (in contrast to segment 5 of Broadhaven and Wexford viruses) was shown to be the major genetic determinant of serotype specificity.


[Study on synthesis and photoluminescence of the complexes of rare earth Eu(III) with 4-vinyl pyridine copolymer].
Rare earth (Eu)-polymer ternary complexes were synthesized using copolymer of 4-vinyl pyridine-methyl acrylate as ligand, and using phen and bipy as coordination ligand. The compositions of the complexes were characterized by FTIR and elemental analysis. The photophysical process of photoluminescence of the complexes were characterized by FTIR and elemental analysis. The photophysical process of photoluminescence of the complexes was discussed more fully by UV spectra and fluorescence spectra. The experiment result shows that the copolymer of 4-vinyl pyridine can coordinate into rare ions directly by nitrogen atom in the pyridine ring. When small ligand reacts with coordination, the fluorescence intensity of the complexes increases greatly because of stronger intermolecular energy transfer.


II.3. Electrograms (ECG, EEG, EMG, EOG).
There is a constant need in medicine to obtain objective measurements of physical and cognitive function as the basis for diagnosis and monitoring of health. The body can be considered as a chemical and electrical system supported by a mechanical structure. Measuring and quantifying such electrical activity provides a means for objective examination of heath status. The term electrogram, from the Greek electro meaning electricity and gram meaning write or record, is the broad definition given to the recording of electrical signal from the body. In order that comparisons of electrical activity can be made against normative data, certain methods and procedures have been defined for different electrograms. This paper reviews these methods and procedures for the more typical electrograms associated with some of the major organs in the body, providing a first point of reference for the reader.


[In vitro proteolysis of casein and gluten by pancreatic enzymes].
The time-course of proteolysis of casein and gluten by pancreatic enzymes (trypsin, chymotrypsin, pancreatic juice) was studied in vitro with various concentrations of enzyme or substrate. The effect of a previous peptic proteolysis, with or without sodium phytate, on proteolysis by pancreatic juice was also studied. Proteolysis was evaluated by measuring the nitrogen in products solubilized in trichloroacetic acid (peptides + amino acids) and in phosphotungstic acid (amino acids). Without a previous peptic proteolysis, at a low enzyme/substrate ratio, the release of peptides and amino acids was greater for casein than for gluten. At a high enzyme/substrate ratio, peptide release by pancreatic juice and chymotrypsin was quite similar for casein and gluten but amino acid release was a little less for gluten. After a previous high peptic proteolysis without sodium phytate, pancreatic juice proteolysis, at a low enzyme/substrate ratio, was increased. Peptide release was similar for casein and gluten but amino acid release was a little less for the gluten. With sodium phytate, the release of peptides was quite similar for casein but was less for gluten. The release of amino acids was diminished in both cases. Thus, gluten was more resistant than casein to the action of pancreatic enzymes. Nevertheless, this was not necessarily observed when those enzymes were used for proteolysis at a high enzyme/substrate ratio or following a high peptic proteolysis. The differences between casein and gluten proteolyses have been discussed according to the specific action of the pancreatic enzymes. In conclusion, in comparing the proteolyses of various foods, it is important to use various enzyme/substrate ratios.


Time-frequency analyses of transient-evoked stimulus-frequency and distortion-product otoacoustic emissions: testing cochlear model predictions.
Time-frequency representations (TFRs) of otoacoustic emissions (OAEs) provide information simultaneously in time and frequency that may be obscured in waveform or spectral analyses. TFRs were applied to transient-evoked stimulus-frequency (SF) and distortion-product (DP) OAEs to test cochlear model predictions. SFOAEs and DPOAEs were elicited in 18 normal-hearing subjects using gated tones and tone pips. Synchronous spontaneous (SS) OAEs were measured to assess their contributions to SFOAEs and DPOAEs. A common form of TFR of measured OAEs was a collection of frequency-specific components often aligned with SSOAE sites, with each component characterized by one or more brief segments or a single long-duration segment. The spectral envelope of evoked OAEs differed from that of the evoking stimulus. Strong emission regions or cochlear "hot spots" were detected, and sometimes accounted for OAE energy observed outside the stimulus bandwidth. Contributions of hot spots and multiple internal reflections to the OAE, and differences between measured and predicted OAE spectra, increased as stimulus level decreased, consistent with level-dependent changes in the estimated cochlear reflectance. Suppression and frequency-pulling effects between components were observed. A recursive formulation was described for the linear coherent reflection emission theory [Zweig and Shera, J. Acoust. Soc. Am. 98, 2018-2047 (1995)] that is well suited for time-domain calculations.


Inhibition of influenza A virus infection in vitro by saliphenylhalamide-loaded porous silicon nanoparticles.
Influenza A viruses (IAVs) cause recurrent epidemics in humans, with serious threat of lethal worldwide pandemics. The occurrence of antiviral-resistant virus strains and the emergence of highly pathogenic influenza viruses have triggered an urgent need to develop new anti-IAV treatments. One compound found to inhibit IAV, and other virus infections, is saliphenylhalamide (SaliPhe). SaliPhe targets host vacuolar-ATPase and inhibits acidification of endosomes, a process needed for productive virus infection. The major obstacle for the further development of SaliPhe as antiviral drug has been its poor solubility. Here, we investigated the possibility to increase SaliPhe solubility by loading the compound in thermally hydrocarbonized porous silicon (THCPSi) nanoparticles. SaliPhe-loaded nanoparticles were further investigated for the ability to inhibit influenza A infection in human retinal pigment epithelium and Madin-Darby canine kidney cells, and we show that upon release from THCPSi, SaliPhe inhibited IAV infection in vitro and reduced the amount of progeny virus in IAV-infected cells. Overall, the PSi-based nanosystem exhibited increased dissolution of the investigated anti-IAV drug SaliPhe and displayed excellent in vitro stability, low cytotoxicity, and remarkable reduction of viral load in the absence of organic solvents. This proof-of-principle study indicates that PSi nanoparticles could be used for efficient delivery of antivirals to infected cells.


[Prevalence of obesity and its relationship with cardiovascular risk in the general population of Albacete].
To find the prevalence of obesity and overweight in the population of Albacete, and their relationship to other cardiovascular and coronary risk factors. A descriptive, crossover population study. Community setting in the province of Albacete. A sample of 2121 people over 18 (843 from the capital and 1278 from the rest of the province) was selected from Albacete's population through stratified random sampling in two stages. Appointments were made with each person by mail. All those attending underwent Anamnesis, a physical examination, ECG and basic analytic tests. 1230 people between 20 and 93 years old were studied. 394 were considered not obese (32%) and 836 were adjudged obese or overweight (68%). 47% of men were overweight and 23.2% of them obese; 33.5% of women were overweight and 32.5% obese. Obesity is a highly prevalent pathology in this province and increases with age. Overweight is more common among men and obesity among women. Hypertension, Hyperlipaemia, Diabetes and high Fibrinogen are more prevalent in obese than in non-obese patients and increase in prevalence along with the obesity. Patients with the risk factors under study are generally obese. The coronary risk calculated was higher among the obese, the risk increasing along with excess of weight.


[History of forensic medical examination service in Belarus in the Soviet period (1918-1991)].
The article presents a part of historical path (1918-1991) of forensic medical examination in the Republic of Belarus, the history of origin, formation and development of forensic medical service and the academical departments of forensic medicine in the BSSR. The creation date of the state forensic medical service on the territory of Belarus can be considered as 3 of July 1918 yr. Since then, the service has passed from the judicial and administrative subdivision of the department of public healthcare of Executive Committee of Vitebsk Council of Workers, Peasant and Red Army Deputies (1918), the department of forensic medicine of the PCH of the BSSR (1919), forensic medical department at the Central Chemical and Biological Station of the PCH of the BSSR (1921) to the Institute of Scientific and Forensic Examination of the BSSR (1929), Central Forensic Medical Laboratory of the PCH of the BSSR (1931) and Republican Bureau of Forensic Medical Examination under the Ministry of Healthcare of the BSSR (1952). The historical stages of the existence of the forensic medical service and its state organizations in the BSSR were following: 1) stage of origin (1918-1922); 2) stage of formation (1921-1941, 1946-1952); 3) stage of development (1952). The main forensic medical experts, who made a significant contribution to the origin, formation and development of forensic medical examination in the territory of Belarus were V.F. Chervakov (1930-1939), S.A. Prilutskiy (1939-1941, 1946-1960), V.K. Steshits (1961-1977) and S.S. Maksimov (from 1977).


Characterization of a binding site for the herpes simplex virus type 1 UL9 origin-binding protein within the UL9 gene.
Gene UL9 of herpes simplex virus type 1 (HSV-1) encodes a sequence-specific origin-binding protein (OBP) that plays a direct and essential role in viral DNA synthesis. A search of the complete HSV-1 genomic sequence for possible OBP binding sites lying outside the known origins of replication revealed the presence of a very close match to the OBP recognition sequence within the UL9 coding region. The ability of OBP to bind to this site (referred to as the 'UL9 box') was confirmed by DNase I footprinting and gel retardation assays, and filter binding experiments demonstrated that the affinity of OBP for the UL9 box was of the same order as for its high affinity sites within the three replication origins. To investigate whether binding of OBP to the UL9 box played a role during viral replication we constructed a mutant virus in which the sequence was altered in such a way as to preserve the encoded amino acid sequence whilst abolishing the ability of OBP to bind. Growth of the virus was indistinguishable from wild-type and no alterations were observed in the accumulation of transcripts from the UL9 region of the genome. In addition, a DNA fragment containing the UL9 box sequence did not exhibit origin activity in a transient assay for viral DNA synthesis. We therefore conclude that binding of OBP to the UL9 box is not essential for virus growth and that expression of the UL9 gene is unlikely to be autoregulated through this site.


A nimodipine-resistant Ca2+ pathway is involved in myogenic tone in a resistance artery.
After stretch, tone develops in ring segments of rabbit ear resistance arteries (75-150 micron unstretched lumen diam). This is myogenic tone, since it is not dependent on the presence of sympathetic nerves, endothelial cells, or the local release of any known vasoactive autacoids. It is sensitive to the extracellular Ca2+ concentration, and disappears in a low Ca2+ (25 microM) environment. Tone is restored immediately on the reintroduction of Ca2+ (0.1-1.6 mM). High K+-induced tone in these resistance arteries is also extracellular Ca2+-dependent. The Ca2+ ED50 of this tone is 2.45 X 10(-4) M, which is significantly lower than that for myogenic tone (Ca2+ ED50; 8.31 X 10(-4) M). Both tone moieties are influenced by inorganic Ca2+ antagonists (Mn2+ and Mg2+) and some organic Ca2+ antagonists (verapamil and diltiazem). Among all the Ca2+ antagonists studied, only Mn2+ completely inhibits myogenic tone. In contrast, myogenic tone is resistant to nimodipine (up to 10(-6) M), the most potent Ca2+ antagonist of K+-induced contraction (ID50; 1.0 X 10(-9) M). Other 1,4-dihydropyridine Ca2+ antagonists such as nifedipine and (-)-PN 200-110 selectively antagonize K+-induced contractions, whereas they do not affect the development and the maintenance of myogenic tone. The fact that the (+)-enantiomer of PN 200-110 does not have an inhibitory effect proves that the 1,4-DHP site is stereospecific. These results indicate that extracellular Ca2+ is essential for both stretch-dependent (myogenic) and K+-induced tone, and the Ca2+ entry pathways for the two tone moieties are differently influenced by dihydropyridines.


Genetic relatedness among human rotaviruses as determined by RNA hybridization.
Viral RNAs from human rotaviruses were compared by gel electrophoresis and by hybridization to probes prepared by in vitro transcription of two well-characterized laboratory strains (Wa and DS-1). Also, the viral RNAs were compared by hybridization to probes prepared from three of the test viruses. Thirteen specimens (diarrheal stools) were obtained from infants and children 5 to 21 months old on a single day at the emergency ward of the Caracas Children's Hospital, and an additional specimen was obtained from the same hospital 6 months before. When the electrophoresed viral RNAs were stained with ethidium bromide and examined by UV light, five different migration patterns (electropherotypes) were distinguished on the basis of differences in mobility of the RNA segments. The hybridization technique that was employed permitted only qualitative comparisons of corresponding genes of different human rotaviruses. Ten of the specimens contained enough virus to yield sufficient RNA for hybridization studies. Eight of the viruses studied by hybridization contained 4 to 11 genes that reacted specifically with the Wa probe to yield double-stranded RNA segments with a mobility similar to that of Wa viral RNA or test virus RNA. The other two viruses contained 11 genes that reacted specifically with the DS-1 hybridization probe to yield double-stranded RNA segments with a mobility similar to DS-1 viral RNA or test virus RNA. A more complex picture emerged when hybridization probes were prepared from three of the test viruses and used to compare the different electropherotypes. Corresponding genes that exhibited similar migration did not necessarily exhibit homology when studied by hybridization. Also, some corresponding genes that exhibited homology did not have the same mobility by gel electrophoresis.


Differential association of modifiable health behaviors with hot flashes in perimenopausal and postmenopausal women.
To determine the association of modifiable factors, such as smoking, body mass index, and alcohol use, with hot flashes, and to ascertain whether the association with hot flashes varies by menopausal stage. A written survey completed by perimenopausal and postmenopausal women enrolling in a randomized, controlled trial of a menopause risk management program in 1999. Survey items included questions on demographics, health status, and health behaviors. A Massachusetts-based health maintenance organization. Female members, age 40 to 65, excluding women with chronic conditions precluding study participation, were randomly selected from an automated medical record system. The majority of the 287 postmenopausal and 468 perimenopausal women participating in the study were white, college educated, and nonsmoking. Approximately 30% of both groups reported experiencing hot flashes. Separate multivariable logistic regression models were developed for perimenopausal and postmenopausal women to identify correlates of reporting any versus no hot flashes. After controlling for age, race, oral contraceptive use, hormone replacement therapy use, and depression, correlates of hot flashes in perimenopausal women were body mass index >/=25 kg/m(2) (odds ration [OR], 2.00; 95% confidence interval [CI], 1.28 to 3.12) and alcohol use of 1 to 5 drinks per week (OR, 0.52; 95% CI, 0.31 to 0.86). The only significant correlate of hot flashes in the postmenopausal population was high dietary fat intake (OR, 0.35; 95% CI, 0.15 to 0.81). Although study respondents were from similiar sociodemographic groups and received their health care in the same health maintenance organization, modifiable factors associated with hot flashes were different for perimenopausal and postmenopausal women.


Use of force plate analysis to assess the analgesic effects of etodolac in horses with navicular syndrome.
To evaluate the musculoskeletal analgesic effect of etodolac administered PO every 12 or 24 hours in chronically lame horses by use of force plate analysis. 22 horses with navicular syndrome. Horses received etodolac (23 mg/kg, PO, q 12 h; n = 7), etodolac (23 mg/kg, PO, q 24 h; 8), or corn syrup (20 mL, PO, q 24 h; control treatment; 7) for 3 days. Combined forelimb peak vertical ground reaction force (PVF) was measured via force plate analysis before the first treatment (baseline) and at 6, 12, 24, and 36 hours after the last treatment. Differences in mean PVF (mPVF) between baseline and subsequent measurements were analyzed (repeated-measures ANOVA) and evaluated for treatment and time effects and treatment-time interaction. Once- or twice-daily administration of etodolac resulted in significant increases in mPVF from baseline at 6, 12, and 24 hours after the last treatment, compared with the control treatment. There were no significant differences in mPVF between the etodolac treatment groups at any time point. In both etodolac treatment groups, there was a significant increase in mPVF from baseline at 6, 12, and 24 hours, compared with that at 36 hours. Etodolac-associated adverse effects were not detected. In horses with navicular syndrome, once-daily oral administration of 23 mg of etodolac/kg appears to provide effective analgesia for as long as 24 hours. Twice-daily administration of etodolac at this same dose does not appear to provide any additional analgesic efficacy or duration of effect.


Preoperative Diagnosis of Tubular Adenoma of Breast - 10 years of experience.
Tubular adenomas are rare benign neoplasms of female breast affecting women, predominantly of child-bearing age group. Definitive diagnosis of this tumor is usually achieved after histopathological study. Clinical, radiological, and even cytological features are often insufficient for accurate diagnosis. The aim of the present study was planned to analyze clinical and radiological features of histologically confirmed cases of tubular adenoma of breast to find out a clue for accurate preoperative cytological diagnosis. In our 10-year research program, all histologically confirmed tubular adenoma cases of breast were studied. Clinico-radio-cytological features, if available, were analyzed and tabulated. Thirty-three cases of tubular adenoma of breast were studied. Radiological and cytological assessments are available for 12 and 26 cases, respectively. Tubular adenoma could be diagnosed in only two cases (7.7%) by cytological assessment, but in none by radiological assessment. Clinico-cyto-radiological assessment could identify the benign nature of the tumors in most cases, but final confirmation was possible only after histopathological study.


Temporal and spatial activation of caspase-like enzymes induced by self-incompatibility in Papaver pollen.
Caspase-like proteases are universal mediators of programmed cell death (PCD). Because plants have no caspase homologs, establishing the nature of their caspase-like activities is of considerable importance to our understanding of PCD in plants. Caspase-3, displaying DEVD specificity, is a key executioner caspase in animal cells. Self-incompatibility (SI) is an important mechanism to prevent self-fertilization and inbreeding in higher plants by inhibiting incompatible pollen. In Papaver rhoeas, SI activates a caspase-3-like/DEVDase activity in incompatible pollen that plays a pivotal role in regulating PCD. Here we characterize the SI-induced caspase-like activities in detail; our work provides insights into the temporal and spatial activation of plant caspase-like enzymes. We show that SI also activates a VEIDase and a LEVDase and that the VEIDase plays a role in SI-induced PCD. The DEVDase and VEIDase are activated remarkably rapidly: detectable within 1-2 h after SI induction; the LEVDase activity peaks later. Importantly, we show live-cell imaging of a DEVDase activity in a higher plant cell; the SI-activated DEVDase has a cytosolic and nuclear localization. We also demonstrate that SI induces a rapid and substantial cytosolic acidification that matches the in vitro pH optima for the SI-induced caspase activities. Because both cytosolic acidification and nuclear caspase localization are observed during apoptosis in animal cells, our data provide striking parallels between SI-induced PCD and apoptosis in animal cells.