Simultaneous detection of multiple genetic aberrations in single cells by spectral fluorescence in situ hybridization.
Spectral fluorescence in situ hybridization (S-FISH) is a novel molecular cytogenetic approach that detects multiple disease-specific chromosomal aberrations in interphase nuclei using combinatorial fluorescence and digital imaging microscopy. A panel of six centromeric probes for chromosomes 7, 8, 9, 10, X, and Y, using a unique two-dye combination of four fluorophores, was developed to assess ploidy in breast tumors, bladder washings, and leukemia. Validation of S-FISH was performed by classic cytogenetics when metaphases were available or by standard fluorescence in situ hybridization (FISH) analyses. S-FISH identified clonal aberrations in newly diagnosed breast tumors and recurrent bladder cancer and revealed minimal residual disease in hyperdiploid acute lymphocytic leukemia, providing "proof of concept." Like standard FISH, aberrations were identified in poor growth/no growth specimen at the single cell level; however, S-FISH provided increased sensitivity over standard FISH by surveying six genetic targets instead of one or two. Disadvantages of the current assay include labor intensive screening and interpretative challenges with signal overlap in highly aneuploid samples and focal plane distortions. S-FISH appears to be a sensitive oncology assay with significant clinical application for early detection of new or reemerging clones, allowing for earlier therapeutic intervention and development of probe panels for individualized therapy.


Radiotherapy versus follow-up in the treatment of pathological stage Ia and Ib non-small cell lung cancer. Early stopped analysis of a randomized controlled study.
This is an analysis of a randomized controlled clinical trial planned to evaluate the effects of adjuvant radiotherapy (AR) on the local recurrence rate in patients with non-small cell lung cancer (NSCLC) with pathological stage (pStage) Ia (pT1N0) and Ib (pT2N0). The effects of AR on the long-term survival have also been marginally evaluated. This clinical trial was planned with the hypothesis that AR on pStage Ia and Ib, R0 NSCLCs was effective on local recurrence rate. From July 1989 through March 1997, 104 patients with NSCLC who presented with pStage Ia and Ib have been observed and treated and entered the study. Male/female ratio was 91:13; the mean age was 62 years (range 41-75 years). All patients underwent major pulmonary resection and homolateral standard hilar and mediastinal lymph node dissection. pStage was T1N0 in 29 and T2N0 in 75 cases. Patients have been randomized 'by chance' into two groups (G1 and G2). G1 received radiotherapy, G2 did not receive any adjuvant treatment. Fifty-two patients entered G1 and 52 entered G2. Post-operative mortality was nil. Seven patients have been excluded from the study (four in G1 and three in G2), due to incomplete follow-up data. We do not report any radiotherapy-related complication or deterioration of lung function. The treatment effect on the local recurrence rate demonstrated a clearly significant protective effect of the AR. No statistically significant difference was found from the comparison of the 5-year survival rate of the treated (83%) versus untreated (70%) patients. No detrimental effect of the radiotherapy has been assessed. AR in the treatment of pStage Ia and Ib NSCLC has been well tolerated and had a significant relative effect on the local recurrence rate but did not significantly modify overall survival even if a positive trend in the group of treated patients is reported.


Telemedicine and eHealth in Poland from 1995 to 2015.
The aim of this study is to present a review based on the literature and proceedings from selected telemedicine conferences. The review was developed using the PRISMA framework. The Embase and PubMed (updated until July 13, 2015) literature databases were searched for telemedicine-related terms and Poland. The literature search identified 129 eligible articles in the databases and 85 in conference proceedings until July 2015. Articles measured as a number of contributions per year presented a similar rising, fluctuating and almost parallel pattern. Fifty-nine percent of the reviewed papers were published in impacted journals. Almost half of all publications presented original papers. The published articles concerned mostly cardiology (16%), family medicine (15%) and pathology (11%). Conference proceedings papers concerned orthopedics (29%, significantly more frequent; p < 0.001) and cardiology (14%). Scientific activity of researchers and practitioners in Poland in the field of telemedicine is not high, but it is increasing over time. There is a tendency to present the research rather in high-quality journals instead of conferences before publication. The occurrence of individual medical specialty telemedicine in Poland may reflect country-specific needs.


Clinically insignificant residual fragments after percutaneous nephrolithotomy: medium-term follow-up.
Clinically insignificant residual fragments (CIRFs), defined as asymptomatic, noninfectious, ≤4 mm fragments, are sometimes observed after percutaneous nephrolithotomy (PCNL). Because the natural history of these fragments is unclear, we investigated the medium-term outcome of these fragments. During a 3-year period, 430 patients underwent PCNL. Overall stone-free rate was 74.5%, and CIRFs were encountered in 22% of cases 3 months after surgery. A total of 38 patients who had CIRFs immediately after PCNL with at least 24 months of follow-up were included in the study. All patients were subjected to periodic follow-up with detailed history, clinical examination, and radiographic follow-up. Serum biochemistry together with urine metabolic evaluation was also performed. The median follow-up was 28.4±5.3 months (range 24-38 mos). Ten (26.3%) patients had a symptomatic episode that necessitated medical therapy during follow-up while others remained asymptomatic. Radiologic assessment showed an increase in the size of the fragments in 8 (21.1%) patients, while the size of the fragments was stable or decreased in 27 (71.1%) cases. Three (7.9%) patients had spontaneous stone passage. Metabolic evaluation revealed abnormalities in 10 (26.3%) patients. Stone analysis revealed magnesium ammonium phosphate in three of eight patients who had an increase in residual fragment size. Also, only two of these eight patients had a metabolic abnormality (one hypocitraturia and one hypercalciuria). Medium-term follow-up of CIRFs after PCNL revealed that progression within 2 years is relatively common. Increase in fragment size is common in patients with struvite stones, and presence of risk factors on 24-hour urine metabolic analysis does not seem to predict growth of observed fragments.


Editing a scientific journal on communication disorders in South Africa: a unique challenge.
The aim of this article is to describe the challenge of editing a scientific journal in the field of communication disorders in the South African context. An overview of the development of the journal and the current editorial policy is described. The distinctiveness and uniqueness of the journal is highlighted. A discussion of the problems encountered and the challenges posed by the future is supplied. In conclusion the question is posed of whether a scientific publication of this nature is relevant and justifiable. It is concluded that such a publication is indeed important and has the right of existence.


The Distally Based Dorsal Metatarsal Artery Perforator Flap: Vascular Study and Clinical Implications.
Background Intrinsic flaps based on the dorsal metacarpal arteries are useful for coverage of dorsal hand, finger, and thumb defects. The purpose of this study was to explore the anatomy of the dorsal metatarsal arteries (DMtAs) in the foot to help define their clinical utility. We observed the size and numbers of distal perforators from the DMtAs and quantified the vascular perfusion pattern of the DMtA perforator across the skin. Methods Ten fresh cadaver feet were injected with latex and dissected to assess the size and number of distal perforators from the DMtAs. Five DMtA perforator flaps were injected with methylene blue to visualize and quantify the vascular territory of the skin flap to understand the clinical possibilities. In addition, a clinical case is described and shown. Results Ten fresh cadaver feet were dissected. The first DMtA was absent in two specimens and the second, third, or fourth DMtA was absent in one specimen each. The available DMtAs had between two and five cutaneous perforators supplying the skin (average, 3.7 perforators per DMtA). The largest perforators to the skin were always seen in the distal half of the DMtA and ranged from 0.4 to 0.8 mm (average, 0.5 mm). Methylene blue injections showed an average flap surface of 21.6 × 47.6 mm. Conclusion This cadaveric study demonstrates the usefulness of the DMtA perforator flap. The flap is a valuable addition to the arsenal of flaps to cover the dorsum of the toe, webspace, or defects exposing tendons on the distal dorsum of the foot.


Spatial heterogeneity may form an inverse camel shaped Arnol'd tongue in parametrically forced oscillations.
Frequency locking in forced oscillatory systems typically organizes in "V"-shaped domains in the plane spanned by the forcing frequency and amplitude, the so-called Arnol'd tongues. Here, we show that if the medium is spatially extended and monotonically heterogeneous, e.g., through spatially dependent natural frequency, the resonance tongues can also display "U" and "W" shapes; we refer to the latter as an "inverse camel" shape. We study the generic forced complex Ginzburg-Landau equation for damped oscillations under parametric forcing and, using linear stability analysis and numerical simulations, uncover the mechanisms that lead to these distinct resonance shapes. Additionally, we study the effects of discretization by exploring frequency locking of oscillator chains. Since we study a normal-form equation, the results are model-independent near the onset of oscillations and, therefore, applicable to inherently heterogeneous systems in general, such as the cochlea. The results are also applicable to controlling technological performances in various contexts, such as arrays of mechanical resonators, catalytic surface reactions, and nonlinear optics.


Systems biology model databases and resources.
Systems biology aims at a quantitative understanding of systemic behaviour as a function of its components and their interactions. In systems biology studies computer models play an important role: (i) to integrate the components' behaviour and (ii) to analyse experimental data sets. With the growing number of kinetic models that are being constructed for parts of biological systems, it has become important to store these models and make them available in a standard form, such that these models can be combined, eventually leading to a model of a complete system. In the present chapter we describe database initiatives that contain kinetic models for biological systems, together with a number of other systems biology resources related to kinetic modelling.


The NAD+ metabolism of Leishmania, notably the enzyme nicotinamidase involved in NAD+ salvage, offers prospects for development of anti-parasite chemotherapy.
NAD+ plays multiple, essential roles in the cell. As a cofactor in many redox reactions it is key in the cellular energy metabolism and as a substrate it participates in many reactions leading to a variety of covalent modifications of enzymes with major roles in regulation of expression and metabolism. Cells may have the ability to produce this metabolite either via alternative de novo synthesis pathways and/or by different salvage pathways. In this issue of Molecular Microbiology, Gazanion et al. (2011) demonstrate that Leishmania species can only rely on the salvage of NAD+ building blocks. One of the enzymes involved, nicotinamidase, is absent from human cells. The enzyme is important for growth of Leishmania infantum and essential for establishing an infection. The crystal structure of the parasite protein has been solved and shows prospects for design of inhibitors to be used as leads for development of new drugs. Indeed, NAD+ metabolism is currently being considered as a promising drug target in various diseases and the vulnerability of Leishmania for interference of this metabolism has been proved in previous work by the same group, by showing that administration of NAD+ precursors has detrimental effect on the pathogenic, amastigote stage of this parasite.


Modified EASIX predicts severe cytokine release syndrome and neurotoxicity after chimeric antigen receptor T cells.
Patients who develop chimeric antigen receptor (CAR) T-cell-related severe cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) exhibit hemodynamic instability and endothelial activation. The EASIX (Endothelial Activation and Stress Index) score (lactate dehydrogenase [LDH; U/L] × creatinine [mg/dL]/platelets [PLTs; 109 cells/L]) is a marker of endothelial damage that correlates with outcomes in allogeneic hematopoietic cell transplantation. Elevated LDH and low PLTs have been associated with severe CRS and ICANS, as has C-reactive protein (CRP), while increased creatinine is seen only in a minority of advanced severe CRS cases. We hypothesized that EASIX and 2 new modified EASIX formulas (simplified EASIX, which excludes creatinine, and modified EASIX [m-EASIX], which replaces creatinine with CRP [mg/dL]), calculated peri-CAR T-cell infusion, would be associated with development of severe (grade ≥ 3) CRS and ICANS. We included 118 adults, 53 with B-acute lymphoblastic leukemia treated with 1928z CAR T cells (NCT01044069) and 65 with diffuse large B-cell lymphoma treated with axicabtagene ciloleucel or tisagenlecleucel. The 3 formulas showed similar predictive power for severe CRS and ICANS. However, low PLTs and high CRP values were the only variables individually correlated with these toxicities. Moreover, only m-EASIX was a significant predictor of disease response. m-EASIX could discriminate patients who subsequently developed severe CRS preceding the onset of severe symptoms (area under the curve [AUC] at lymphodepletion, 80.4%; at day -1, 73.0%; and at day +1, 75.4%). At day +3, it also had high discriminatory ability for severe ICANS (AUC, 73%). We propose m-EASIX as a clinical tool to potentially guide individualized management of patients at higher risk for severe CAR T-cell-related toxicities.


Coupling of neural activity to blood flow in olfactory glomeruli is mediated by astrocytic pathways.
Functional neuroimaging uses activity-dependent changes in cerebral blood flow to map brain activity, but the contributions of presynaptic and postsynaptic activity are incompletely understood, as are the underlying cellular pathways. Using intravital multiphoton microscopy, we measured presynaptic activity, postsynaptic neuronal and astrocytic calcium responses, and erythrocyte velocity and flux in olfactory glomeruli during odor stimulation in mice. Odor-evoked functional hyperemia in glomerular capillaries was highly correlated with glutamate release, but did not require local postsynaptic activity. Odor stimulation induced calcium transients in astrocyte endfeet and an associated dilation of upstream arterioles. Calcium elevations in astrocytes and functional hyperemia depended on astrocytic metabotropic glutamate receptor 5 and cyclooxygenase activation. Astrocytic glutamate transporters also contributed to functional hyperemia through mechanisms independent of calcium rises and cyclooxygenase activation. These local pathways initiated by glutamate account for a large part of the coupling between synaptic activity and functional hyperemia in the olfactory bulb.


The difficult concussion patient: what is the best approach to investigation and management of persistent (>10 days) postconcussive symptoms?
Concussion in sport typically recovers clinically within 10 days of injury. In some cases, however, symptoms may be prolonged or complications may develop. The objectives of the current paper are to review the literature regarding the difficult concussion and to provide recommendations for an approach to the investigation and management of patients with persistent symptoms. A qualitative review of the literature on concussion in sport was conducted with a focus on prolonged recovery, long-term complications and management including investigation and treatment strategies. MEDLINE and Sports Discus databases were reviewed. Persistent symptoms (>10 days) are generally reported in 10-15% of concussions. This figure may be higher in certain sports (eg, ice hockey) and populations (eg, children). In general, symptoms are not specific to concussion and it is important to consider and manage coexistent pathologies. Investigations may include formal neuropsychological testing and conventional neuroimaging to exclude structural pathology. Currently, there is insufficient evidence to recommend routine clinical use of advanced neuroimaging techniques or genetics markers. Preliminary studies demonstrate the potential benefit of subsymptom threshold activity as part of a comprehensive rehabilitation programme. Limited research is available on pharmacological interventions. Cases of concussion in sport where clinical recovery falls outside the expected window (ie, 10 days) should be managed in a multidisciplinary manner by healthcare providers with experience in sports-related concussion. Important components of management, after the initial period of physical and cognitive rest, include associated therapies such as cognitive, vestibular, physical and psychological therapy, assessment for other causes of prolonged symptoms and consideration of a graded exercise programme at a level that does not exacerbate symptoms.


A novel technique for the examination of skin biopsies by laser capture microdissection.
Skin is an inherently heterogeneous tissue, thus the procurement of pure cell populations is critical for the accurate correlation of a molecular profile to a particular cell type or histological location. Laser Capture Microdissection (LCM) permits the efficient procurement of cells and mapping of genetic changes from histologically prepared samples. This paper describes a robust LCM protocol established in our laboratory for the extraction of high quality DNA which sequenced from 100% of microdissected samples without the need for cloning. The unique properties of skin, in particular its strong intercellular adhesive forces, have dictated a significant modification to the normal procedure of tissue preparation to ensure reliable cell procurement. Using the methods outlined below we were able to precisely map the pattern of genomic mutations in our target gene of interest in normal skin, actinic keratosis and squamous cell carcinoma. The capability to select pure cell populations from the skin will revolutionise our ability to understand the processes involved in cutaneous tumourigenesis.


Bullous pemphigoid:therapy in patients with and without diabetes mellitus.
Retrospective analysis of 34 patients with bullous pemphigoid (BP) showed 24 (70.6%) with widespread disease and 10 (29.4%) with localized disease. Initial prednisone therapy was significantly lower for patients with localized BP, mean dose of 38 mg vs 58 mg, compared with widespread disease. Fifteen of 24 (62%) patients responded; however, 9 (38%) failed to respond to moderate-dose prednisone therapy and ultimately required daily doses of 100 mg or more and/or an immunosuppressive agent. Adult-onset diabetes mellitus was present in 14 of 34 (41%) patients. Initial prednisone dose for diabetic patients with widespread BP was significantly greater, 70 mg vs 54 mg, compared with nondiabetic patients. Addition of methotrexate, 2.5 to 15 mg, every other day or twice-weekly oral administration, was effective when moderate-dose prednisone therapy failed. It is possible that insulin-dependent diabetic patients with BP should be preferentially started on immunosuppressive drugs.


Examining the Impact of Digital Components Across Different Phases of Treatment in a Blended Care Cognitive Behavioral Therapy Intervention for Depression and Anxiety: Pragmatic Retrospective Study.
Depression and anxiety incur significant personal and societal costs. Effective psychotherapies exist, such as cognitive behavioral therapy (CBT); however, timely access to quality care is limited by myriad barriers. Blended care therapy models incorporate traditional face-to-face therapy with scalable, digital components of care, expanding the reach of evidence-based care. The aim of this study is to determine the effectiveness of a blended care CBT program (BC-CBT) in real-world settings and examine the unique impacts of the (1) digital components of care (video lessons and digital exercises) and (2) phase of treatment (early versus late) in decreasing symptoms of anxiety and depression. This retrospective cohort analysis included 3401 US-based individuals enrolled in a BC-CBT program, who presented with clinical levels of depression and/or anxiety. The treatment program consisted of regular therapy sessions augmented by clinician-assigned digital video lessons and exercises. A growth curve model incorporating time-varying covariates examined the relationship between engagement with BCT components (ie, therapy sessions, digital video lessons, and digital exercises) during the early (weeks 0-7) and late (weeks 8-15) phases of treatment, and weekly symptom reports on depression and anxiety measures. On average, a significant decline in depression and anxiety symptoms was observed during the initial weeks of treatment (P<.001), with a continued, though slower, decline over subsequent weeks (P<.001). Each session completed was associated with significant decreases in anxiety (b=-0.72) and depression (b=-0.83) in the early phase, as well as in the late phase (anxiety, b=-0.47; depression, b=-0.27). Significant decreases in anxiety (b=-0.15) and depression (b=-0.12) were observed for time spent on video lessons (measured in 10-minute intervals) in the early phase of treatment. Engaging with exercises was associated with statistically significant increases in anxiety symptoms (b=0.03) during the early phase of treatment. However, sensitivity analyses examining the effects of exercises in isolation revealed significant decreases in anxiety (b=-0.05) in the early phase, suggesting a potential suppression effect in the larger model. Using a retrospective cohort design, therapy sessions and digital video lessons were uniquely predictive of improvements in depression and anxiety symptoms, and their effects were modulated based on the phase of treatment (early vs late). Future research should investigate whether other treatment variables, such as therapeutic alliance or familiarity with technology, are related to differential effects on various components of care.


Blastochloris gulmargensis sp. nov., isolated from an epilithic phototrophic biofilm.
A novel Gram-negative, motile, bacteriochlorophyll b-containing purple non-sulfur bacterium, strain JA248(T), was isolated from phototrophic enrichments of a yellow-green epilithic biofilm sample collected from Gulmarg, India. The genomic DNA G+C content of strain JA248(T) was 63.8 mol%. A phylogenetic tree based on 16S rRNA gene sequence analysis showed that strain JA248(T) had highest similarity to members of the genus Blastochloris and was closely related to Blastochloris sulfoviridis DSM 729(T) (98.5 % sequence similarity) and Blastochloris viridis DSM 133(T) (98.4 %) of the class Alphaproteobacteria. Strain JA248(T) was characterized based on polyphasic taxonomy, and distinct phenotypic and molecular differences based on DNA-DNA hybridization (relatedness of <46.5 % with the two species of the genus Blastochloris), multilocus sequence analysis, and phenotypic and chemotaxonomic evidence separated strain JA248(T) from other species of the genus Blastochloris. Strain JA248(T) therefore represents a novel species in the genus Blastochloris, for which the name Blastochloris gulmargensis sp. nov. is proposed. The type strain is JA248(T) ( = JCM 14795(T) = DSM 19786(T)).


Seizure outcomes after resective surgery for extra-temporal lobe epilepsy in pediatric patients.
While temporal lobe epilepsy (TLE) is the most common epilepsy syndrome in adults, seizures in children are more often extratemporal in origin. Extra-temporal lobe epilepsy (ETLE) in pediatric patients is often medically refractory, leading to significantly diminished quality of life. Seizure outcomes after resective surgery for pediatric ETLE vary tremendously in the literature, given diverse patient and epilepsy characteristics and small sample sizes. The authors performed a systematic review and meta-analysis of studies including 10 or more pediatric patients (age ≤ 19 years) published over the last 20 years examining seizure outcomes after resective surgery for ETLE, excluding hemispherectomy. Thirty-six studies were examined. These 36 studies included 1259 pediatric patients who underwent resective surgery for ETLE. Seizure freedom (Engel Class I outcome) was achieved in 704 (56%) of these 1259 patients postoperatively, and 555 patients (44%) continued to have seizures (Engel Class II-IV outcome). Shorter epilepsy duration (≤ 7 years, the median value in this study) was more predictive of seizure freedom than longer (> 7 years) seizure history (odds ratio [OR] 1.52, 95% confidence interval [CI] 1.07-2.14), suggesting that earlier intervention may be beneficial. Also, lesional epilepsy was associated with better seizure outcomes than nonlesional epilepsy (OR 1.34, 95% CI 1.19-1.49). Other predictors of seizure freedom included an absence of generalized seizures (OR 1.61, 95% CI 1.18-2.35) and localizing ictal electroencephalographic findings (OR 1.55, 95% CI 1.24-1.93). In conclusion, seizure outcomes after resective surgery for pediatric ETLE are less favorable than those associated with temporal lobectomy, but seizure freedom may be more common with earlier intervention and lesional epilepsy etiology. Children with continued debilitating seizures despite failure of multiple medication trials should be referred to a comprehensive pediatric epilepsy center for further medical and surgical evaluation.


Biocapacity optimization in regional planning.
Ecological overshoot has been accelerating across the globe. Optimizing biocapacity has become a key to resolve the overshoot of ecological demand in regional sustainable development. However, most literature has focused on reducing ecological footprint but ignores the potential of spatial optimization of biocapacity through regional planning of land use. Here we develop a spatial probability model and present four scenarios for optimizing biocapacity of a river basin in Northwest China. The potential of enhanced biocapacity and its effects on ecological overshoot and water consumption in the region were explored. Two scenarios with no restrictions on croplands and water use reduced the overshoot by 29 to 53%, and another two scenarios which do not allow croplands and water use to increase worsened the overshoot by 11 to 15%. More spatially flexible transition rules of land use led to higher magnitude of change after optimization. However, biocapacity optimization required a large amount of additional water resources, casting considerable pressure on the already water-scarce socio-ecological system. Our results highlight the potential for policy makers to manage/optimize regional land use which addresses ecological overshoot. Investigation on the feasibility of such spatial optimization complies with the forward-looking policies for sustainable development and deserves further attention.


Unveiling the Effect of Low pH on the SARS-CoV-2 Main Protease by Molecular Dynamics Simulations.
(1) Background: Main Protease (Mpro) is an attractive therapeutic target that acts in the replication and transcription of the SARS-CoV-2 coronavirus. Mpro is rich in residues exposed to protonation/deprotonation changes which could affect its enzymatic function. This work aimed to explore the effect of the protonation/deprotonation states of Mpro at different pHs using computational techniques. (2) Methods: The different distribution charges were obtained in all the evaluated pHs by the Semi-Grand Canonical Monte Carlo (SGCMC) method. A set of Molecular Dynamics (MD) simulations was performed to consider the different protonation/deprotonation during 250 ns, verifying the structural stability of Mpro at different pHs. (3) Results: The present findings demonstrate that active site residues and residues that allow Mpro dimerisation was not affected by pH changes. However, Mpro substrate-binding residues were altered at low pHs, allowing the increased pocket volume. Additionally, the results of the solvent distribution around Sγ, Hγ, Nδ1 and Hδ1 atoms of the catalytic residues Cys145 and His41 showed a low and high-water affinity at acidic pH, respectively. It which could be crucial in the catalytic mechanism of SARS-CoV-2 Mpro at low pHs. Moreover, we analysed the docking interactions of PF-00835231 from Pfizer in the preclinical phase, which shows excellent affinity with the Mpro at different pHs. (4) Conclusion: Overall, these findings indicate that SARS-CoV-2 Mpro is highly stable at acidic pH conditions, and this inhibitor could have a desirable function at this condition.


Effect of sorghum grain variety and reconstitution on site and extent of starch and protein digestion in steers.
Hetero-yellow (HY), red (RED) and brown (BR, high tannin) sorghums were fed dry-rolled or reconstituted (RED and BR only) to evaluate the effect of variety and reconstitution on the site and extent of starch and protein digestion in steers fitted with ruminal, duodenal and ileal cannulae. Processed grains were incorporated into 88% sorghum (DM basis) diets fed at 2% of body weight in a 5 X 5 Latin square. Ruminal fermentation of organic matter, starch and protein tended to be lower for the dry-rolled RED than for either the dry-rolled HY or BR sorghum. Digestion of organic matter (OM) and starch in the small intestine was very low for dry-rolled sorghums. Total tract starch digestibility was lower for the dry-rolled RED sorghum (86.9%) than the BR (90.8%) and HY (91.4%). Nitrogen (N) digestibility ranged from 53.1% for the dry-rolled BR to 64.5% for the HY. Tannins were extensively (95.2%) degraded in the rumen, which may have enhanced fermentation of the BR sorghum. Reconstitution increased (P less than .05) total-tract starch digestion of the RED and tended to increase starch digestion of the BR as well. Total N flow to the duodenum tended to increase with reconstitution, with most of the increase being due to greater (P less than .05) microbial-N. Reconstitution also increased (P less than .05) total-tract N digestibility of the RED. The response to reconstitution for the RED sorghum appeared to be due primarily to an increase (P less than .10) in the extent of fermentation of organic matter and starch in the rumen. Reconstitution of BR, however, enhanced disappearance of starch from the small intestine. In both cases, most (97.3%) of the digestible starch of the reconstituted sorghums had disappeared before the terminal ileum. In contrast, 14.5% (621 g) of the digestible starch of dry-rolled RED disappeared in the large intestine. Sorghum grain variety and reconstitution appear to alter site and extent of starch and protein digestion, which may result in variable performance of cattle fed sorghum grain diets.


Using "spinal shrinkage" as a trigger for motivating students to learn about obesity and adopt a healthy lifestyle.
Obesity is a global problem; however, relatively little attention is directed toward preparing and inspiring students of medicine and allied medical sciences to address this serious matter. Students are not routinely exposed to the assessment methods for obesity, its overall prevalence, causative factors, short- and long-term consequences, and its management by lifestyle modification. This physiology laboratory exercise involving students of medicine (n = 106) was developed to 1) introduce medical students to methods of obesity assessment and to differentiate between general and abdominal obesity, 2) generate an interest and sensitivity about obesity, and 3) stimulate thinking about modification of their lifestyle in relation to eating habits, weight control, and physical activity. Spinal shrinkage (the difference between the standing height of a person and his/her recumbent length) was used as an immediate observable parameter to demonstrate the effect of adiposity. Spinal shrinkage is recognized as an index of the compressive forces acting on the spine and is related to body mass index. A positive correlation (r = 0.365, P < 0.05) was observed between body mass index and spinal shrinkage. A questionnaire was used to assess student responses to this exercise. Students were motivated to engage in more physical activity (74%), adopt healthier eating (63%), and enhance their knowledge about obesity (67%). They expressed keen interest in the laboratory exercise and found the sessions enjoyable (91%). The laboratory exercise proved to be a success in motivating the students to actively learn and inquire about obesity and to adopt a healthier lifestyle.


Application of Texture Analysis to Study Small Vessel Disease and Blood-Brain Barrier Integrity.
We evaluate the alternative use of texture analysis for evaluating the role of blood-brain barrier (BBB) in small vessel disease (SVD). We used brain magnetic resonance imaging from 204 stroke patients, acquired before and 20 min after intravenous gadolinium administration. We segmented tissues, white matter hyperintensities (WMH) and applied validated visual scores. We measured textural features in all tissues pre- and post-contrast and used ANCOVA to evaluate the effect of SVD indicators on the pre-/post-contrast change, Kruskal-Wallis for significance between patient groups and linear mixed models for pre-/post-contrast variations in cerebrospinal fluid (CSF) with Fazekas scores. Textural "homogeneity" increase in normal tissues with higher presence of SVD indicators was consistently more overt than in abnormal tissues. Textural "homogeneity" increased with age, basal ganglia perivascular spaces scores (p < 0.01) and SVD scores (p < 0.05) and was significantly higher in hypertensive patients (p < 0.002) and lacunar stroke (p = 0.04). Hypertension (74% patients), WMH load (median = 1.5 ± 1.6% of intracranial volume), and age (mean = 65.6 years, SD = 11.3) predicted the pre/post-contrast change in normal white matter, WMH, and index stroke lesion. CSF signal increased with increasing SVD post-contrast. A consistent general pattern of increasing textural "homogeneity" with increasing SVD and post-contrast change in CSF with increasing WMH suggest that texture analysis may be useful for the study of BBB integrity.


Comparison of five agglutination tests for identification of Staphylococcus aureus.
Various commercially produced agglutination kits are widely used for the identification of Staphylococcus aureus. These kits detect the presence of protein A and/or clumping factor on S. aureus. The literature has shown that methicillin-resistant S. aureus (MRSA) isolates which are deficient in both clumping factor and protein A may be misidentified. Two products, Slidex and Staphaurex Plus, utilize specific anti-S. aureus antibodies, potentially giving them greater sensitivity compared to products without these antibodies. We report a prospective study designed to compare the performance characteristics of Fastaph, Slidex, Staphaurex, Staphaurex Plus, Staphyloslide, and the tube coagulase test for the identification of staphylococcal isolates. All discrepant isolates were tested with the Gen-Probe AccuProbe S. aureus test and were identified to the species level with conventional reference biochemicals. A total of 1,193 isolates were tested, including 33 MRSA and 423 methicillin-sensitive S. aureus isolates. The sensitivities and specificities of the tests, respectively, were as follows: Fastaph, 99.1 and 98.9%; Slidex, 99.6 and 96.4%; Staphaurex, 98.9 and 99.9%; Staphaurex Plus, 99.6 and 93.9%; Staphyloslide, 99.1 and 98.9%; and tube coagulase, 99.3 and 100%. Sensitivity was excellent for all of the products tested. The specificities of Fastaph, Staphaurex, and Staphyloslide were excellent, while Staphaurex Plus and Slidex demonstrated less optimal results.


Regulation of nitrogenase synthesis in intact cells of Rhodospirillum rubrum: inactivation of nitrogen fixation by ammonia, L-glutamine and L-asparagine.
The synthesis of nitrogenase by intact cells of Rhodospirillum rubrum was repressed in N-free media supplemented with L-glutamine or L-asparagine, but was unaffected by the presence of L-glutamate, L-aspartate or L-histidine. Specific activities attained by cultures in supplemented media maintained under Ar-CO2 were 2 to 3 times higher than those in N-free medium under N2-CO2. A loss in total activity occurred both in cultures growing with N2 after maximum activity had been reached, and in cultures maintained under Ar when the gas phase was changed to N2. There was a rapid loss in nitrogen-fixing activity when low concentrations of NH4+, L-glutamine or L-asparagine were added to cultures with high activities, but this could be recovered in the absence of demonstrable protein synthesis. During growth, the degree of inactivation brought about by 0-5 mM-inactivator increased to 80 to 90%, and NH4+ excreted into the medium reached a maximum concentration towards the end of exponential growth.


Coexistence of Antibiotic Resistance Genes and Virulence Factors Deciphered by Large-Scale Complete Genome Analysis.
Widespread use of antibiotics has enhanced the evolution of highly resilient pathogens and poses a severe risk to human health via coselection of antibiotic resistance genes (ARGs) and virulence factors (VFs). In this study, we rigorously evaluate the abundance relationship and physical linkage between ARGs and VFs by performing a comprehensive analysis of 9,070 bacterial genomes isolated from multiple species and hosts. The coexistence of ARGs and VFs was observed in bacteria across distinct phyla, pathogenicities, and habitats, especially among human-associated pathogens. The coexistence patterns of gene elements in different habitats and pathogenicity groups were similar, presumably due to frequent gene transfer. A shorter intergenic distance between mobile genetic elements and ARGs/VFs was detected in human/animal-associated bacteria, indicating a higher transfer potential. Increased accumulation of exogenous ARGs/VFs in human pathogens highlights the importance of gene acquisition in the evolution of human commensal bacteria. Overall, the findings provide insights into the genic features of combinations of ARG-VF and expand our understanding of ARG-VF coexistence in bacteria.IMPORTANCE Antibiotic resistance has become a serious global health concern. Despite numerous case studies, a comprehensive analysis of ARG and VF coexistence in bacteria is lacking. In this study, we explore the coexistence profiles of ARGs and VFs in diverse categories of bacteria by using a high-resolution bioinformatics approach. We also provide compelling evidence of unique ARG-VF gene pairs coexisting in specific bacterial genomes and reveal the potential risk associated with the coexistence of ARGs and VFs in organisms in both clinical settings and environments.


Transnational corporations and oral health: examples from the sugar industry.
This paper seeks to describe the political behavior of transnational corporations (TNCs) related to sugars and dental caries. The paper begins by exploring dental caries as a political issue. It then provides a brief overview of key actors (expanders--e.g. public health advocates working to make policy action on sugar likely, and containers--e.g. TNC's working to prevent policy action on sugar) and the importance of problem definition in public policy making. The paper then compares how expanders and containers frame the problem of sugars and dental caries. Based upon a policy analysis framework, categories used to frame problems include incidence, causality, severity, crisis, characteristics of the problem population, values, and solutions. These categories are discussed with application to debates about public policy solutions to the problem of dental caries. It then concludes by highlighting some tensions that remain in tackling dental caries through legislation and regulation.


Occupational exposure to isoflurane during anaesthesia induction with standard and scavenging double masks in dogs, pigs and ponies.
Induction of anaesthesia using a face mask may cause workplace pollution with anaesthetics. The aim of this study was to compare the effect of the use of a standard versus a scavenging double face mask on isoflurane pollution during induction of anaesthesia in experimental animals: six dogs, 12 pigs and five ponies. Pigs were anaesthetized only once using either mask type randomly (n = 6). Dogs and ponies were anaesthetized twice, using different mask types for each occasion in a random order with at least 14 days between experiments. The masks were attached to a Bain breathing system (dogs and pigs) or to a circle system (ponies) using a fresh gas flow of 300 or 50 mL/kg/min, respectively, with 5% vaporizer dial setting. Isoflurane concentrations were measured in the anaesthetist's breathing zone using an infrared photoacoustic spectrometer. The peak isoflurane concentrations (pollution) during baseline and induction periods were compared with Wilcoxon test in all species, and values between the mask types were compared with either Wilcoxon (ponies and dogs) or Mann-Whitney tests (pigs) (P < 0.05). Pollution was higher during induction when compared with baseline regardless of the mask type used but it was only statistically significant in dogs and pigs. Pollution was lower during induction with double versus single masks but it was only significant in pigs. Despite the lack of statistical significance, large and consistent differences were noted in all species, hence using scavenging masks is recommended to reduce isoflurane workplace pollution.


Exploring the reverse J-shaped curve between urea reduction ratio and mortality.
Although accepted worldwide as valid measures of dialysis adequacy, neither the Kt/V (urea clearance determined by kinetic modeling) nor the urea reduction ratio (URR) have unambiguously predicted survival in hemodialysis patients. Because the ratio Kt/V can be high with either high Kt (clearance x time) or low V (urea volume of distribution) and V may be a proxy for skeletal muscle mass and nutritional health, we hypothesized that the increase in the relative risk of death observed among individuals dialyzed in the top 10 to 20% of URR or Kt/V values might reflect a competing risk of malnutrition. A total of 3,009 patients who underwent bioelectrical impedance analysis were stratified into quintiles of URR. Laboratory indicators of nutritional status and two bioimpedance-derived parameters, phase angle and estimated total body water, were compared across quintiles. The relationship between dialysis dose and mortality was explored, with a focus on how V influenced the structure of the dose-mortality relationship. There were statistically significant differences in all nutritional parameters across quintiles of URR or Kt/V, indicating that patients in the fifth quintile (mean URR, 74.4 +/- 3.1%) were more severely malnourished on average than patients in all or some of the other quintiles. The relationship between URR and mortality was decidedly curvilinear, resembling a reverse J shape that was confirmed by statistical analysis. An adjustment for the influence of V on URR or Kt/V was performed by evaluating the Kt-mortality relationship. There was no evidence of an increase in the relative risk of death among patients treated with high Kt. Higher Kt was associated with a better nutritional status. We conclude that the increase in mortality observed among those patients whose URR or Kt/V are among the top 10 to 20% of patients reflects a deleterious effect of malnutrition (manifest by a reduced V) that overcomes whatever benefit might be derived from an associated increase in urea clearance. Identification of patients who achieve extremely high URR (>75%) or single-pooled Kt/V (>1.6) values using standard dialysis prescriptions should prompt a careful assessment of nutritional status. Confounding by protein-calorie malnutrition may limit the utility of URR or Kt/V as a population-based measure of dialysis dose.


Time to match; when do homologous chromosomes become closer?
In most eukaryotes, pairing of homologous chromosomes is an essential feature of meiosis that ensures homologous recombination and segregation. However, when the pairing process begins, it is still under investigation. Contrasting data exists in Mus musculus, since both leptotene DSB-dependent and preleptotene DSB-independent mechanisms have been described. To unravel this contention, we examined homologous pairing in pre-meiotic and meiotic Mus musculus cells using a three-dimensional fluorescence in situ hybridization-based protocol, which enables the analysis of the entire karyotype using DNA painting probes. Our data establishes in an unambiguously manner that 73.83% of homologous chromosomes are already paired at premeiotic stages (spermatogonia-early preleptotene spermatocytes). The percentage of paired homologous chromosomes increases to 84.60% at mid-preleptotene-zygotene stage, reaching 100% at pachytene stage. Importantly, our results demonstrate a high percentage of homologous pairing observed before the onset of meiosis; this pairing does not occur randomly, as the percentage was higher than that observed in somatic cells (19.47%) and between nonhomologous chromosomes (41.1%). Finally, we have also observed that premeiotic homologous pairing is asynchronous and independent of the chromosome size, GC content, or presence of NOR regions.


Cocaine activates platelets and increases the formation of circulating platelet containing microaggregates in humans.
To determine whether there is evidence of platelet activation following in vivo cocaine administration in humans, as cocaine abuse is associated with myocardial infarction and stroke, and platelet activation leading to thrombosis is a possible mechanism. University hospital. Following a randomised, double blind crossover design, 14 healthy volunteers were studied twice, receiving cocaine (2 mg/kg intranasally) once and placebo once. Flow cytometric analysis of P-selectin expression (an alpha granule membrane protein found on the surface of activated platelets), quantification of the platelet specific proteins platelet factor 4 and beta thromboglobulin, and measurement of platelet containing microaggregate and platelet microparticle (fragment) formation were used to assess platelet activation. Circulating von Willebrand factor antigen (vWF) was measured to evaluate a possible role of endothelial stimulation concurrent with platelet activation. There was an increase in both platelet factor 4 (mean (SD), 16 (7) to 39 (22) IU/ml, p = 0. 04) and beta thromboglobulin (70 (20) to 98 (26) IU/ml, p < 0.01) at 120 minutes following cocaine administration. Platelet containing microaggregate formation was increased at 40 minutes (from 47 (3.2)% to 54 (2.0)%, p < 0.001) and 80 minutes (55 (2.5)%, p = 0.04). Bleeding time decreased following cocaine from 10 (1) to 9 (1) minutes (p = 0.07). No changes in any of the measured variables were noted following placebo administration. Cocaine exposure causes platelet activation, alpha granule release, and platelet containing microaggregate formation. These data support the view that cocaine, even at the relatively low doses commonly self administered by occasional abusers, may promote thrombosis and predispose healthy individuals to ischaemic events. Platelet inhibitors should be considered early in any patient with suspected cocaine related ischaemia.


HLA-DQA1 alleles and the presence of Helicobacter pylori antibodies.
Recent reports suggest that the HLA-DQA1 gene may be important in determining susceptibility to and outcome of Helicobacter pylori infection. To determine if there is an association between HLA-DQA1 alleles and H. pylori antibodies, DQA1 alleles and H. pylori-specific antibodies were determined in 199 random subjects of Finnish origin (mean age 43 years, range 22-69 years). H. pylori-specific class IgG antibodies were measured using the EIA method (Pyloriset-EIA-G, Orion Diagnostica, Espoo, Finland). HLA-DQA1 typing was carried out using PCR-SSP (PCR with sequence-specific primers). There were 64 subjects with H. pylori-specific class IgG antibodies (ab+) and 135 subjects without H. pylori-specific class IgG antibodies (ab-). Gene and phenotype frequencies of HLA-DQA1 alleles were similar in the ab+ and ab- subjects (P = NS). The data suggest that no single HLA-DQA1 allele is associated with the presence of serum antibodies against H. pylori.


Phosphaketenes as building blocks for the synthesis of triphospha heterocycles.
Unsaturated phosphorus compounds, such as phosphaalkenes and phosphaalkynes, show a versatile reactivity in cycloadditions. Although phosphaketenes (R-P=C=O) have been known for three decades, their chemistry has remained limited. Herein, we show that heteroatom-substituted phosphaketenes, R(3) E-P=C=O (E=Si, Sn), are building blocks for silyl- and stannyl-substituted five-membered heterocycles containing three phosphorous atoms. The structure of the heterocyclic anion depends on the nature of the tetrel atom involved. Although the silyl analogue [P(3)C(2) (OSiR(3))(2)](-) is an aromatic 1,2,4-triphospholide, the stannyl compound [P(CO)(2) (PSnR(3))(2)](-) is a 1,2,4-triphosphacyclopenta-3,5-dionate with a delocalized OCPCO fragment. Because of their anionic character, these compounds can easily be used as building blocks, for example, in the preparation of a silyl-functionalized hexaphosphaferrocene or the parent 1,2,4-triphosphacyclopenta-3,5-dionate [P(CO)(2) (PH)(2)](-). NMR spectroscopic investigations and computations have shown that the heterocycle-formation reactions presented herein are remarkably complex.


Ibrutinib enhances the antitumor immune response induced by intratumoral injection of a TLR9 ligand in mouse lymphoma.
We have designed a novel therapeutic approach for lymphoma that combines targeted kinase inhibition with in situ vaccination. Intratumoral injection of an unmethylated cytosine guanine dinucleotide (CpG)-enriched oligodeoxynucleotide, an agonist for the toll-like receptor 9 (TLR9), induces the activation of natural killer cells, macrophages, and antigen presenting cells that control tumor growth at the local site. Ibrutinib, an irreversible inhibitor of Bruton's tyrosine kinase, a key enzyme in the signaling pathway downstream of B-cell receptor, is an effective treatment against many types of B-cell lymphomas. The combination of intratumoral injection of CpG with systemic treatment by ibrutinib resulted in eradication of the tumors not only in the injected site, but also at distant sites. Surprisingly, this combinatorial antitumor effect required an intact T-cell immune system since it did not occur in nude, severe combined immunodeficiency, or T-cell depleted mice. Moreover, T cells from animals treated with intratumoral CpG and ibrutinib prevented the outgrowth of newly injected tumors. This result suggests that ibrutinib can induce immunogenic cell death of lymphoma cells and that concomitant stimulation of antigen-presenting cells in the tumor microenvironment by toll-like receptor ligands can lead to a powerful systemic antitumor immune response.


Liposuction-Induced Fat Embolism Syndrome: A Brief Review and Postmortem Diagnostic Approach.
Lipoplasty, or liposuction, the surgical process of removing excess fat, is an elective procedure with rising frequency in the United States. Fat embolism syndrome is a clinical diagnosis and is defined as fat in the circulation with an identifiable clinical pattern of signs and symptoms (eg, hypoxemia, respiratory insufficiency, neurologic impairment, and petechial rash) that occur in the appropriate clinical context. Fat embolism syndrome following liposuction is a life-threatening complication, although its incidence is low. Currently, there is no specific therapy for fat embolism syndrome, so prevention, early detection, and supportive therapy are critical. Many cases of fat embolism syndrome are undiagnosed or misdiagnosed; however, postmortem examination can provide the means for appropriate diagnosis. Therefore, a pathologist must keep a keen eye, as microscopic fat emboli are difficult to appreciate with routine tissue processing and staining.


Direct Marketing Promotion and Electronic Cigarette Use Among US Adults, National Adult Tobacco Survey, 2013-2014.
The use of electronic cigarettes (e-cigarettes) among US adults has increased since 2007. The objective of this study was to investigate the prevalence of direct marketing promotion of e-cigarettes and its association with e-cigarette use among US adults. We used using data from the 2013-2014 National Adult Tobacco Survey (NATS) to estimate prevalence of e-cigarette promotions received by mail or email. Multinomial logistic regression was used to examine the associations between e-cigarette promotions and the prevalence and frequency of e-cigarette use among US adults. In the 2013-2014 survey period, 7.1% of adults (about 16.0 million) reported receiving mail or email e-cigarette promotions in the previous 6 months; 3.2% received mail promotions, and 5.1% received email promotions. A higher prevalence of promotions was found among men versus women, adults aged under 65 years versus those older, current e-cigarette users, current smokers, and people with no smoking restriction rules in their homes or vehicles. In the multivariable analysis, receiving mail or email e-cigarette promotions was associated with higher odds of being current e-cigarette users (adjusted odds ratio [aOR] = 2.0; P < .001) than being noncurrent e-cigarette users. Receiving promotions was also associated with higher odds of using e-cigarettes some days (aOR = 1.6; P = .006) or every day (aOR = 1.7; P = .008) than using e-cigarettes rarely. Receipt of e-cigarette direct marketing promotions was prevalent among US adults. Receiving e-cigarette promotions was associated with increased odds of both prevalence and frequency of e-cigarette use. Future longitudinal studies are needed to measure causal effects of e-cigarette promotions on e-cigarette use among adults.


Defining the transcriptomic landscape of the developing enteric nervous system and its cellular environment.
Motility and the coordination of moving food through the gastrointestinal tract rely on a complex network of neurons known as the enteric nervous system (ENS). Despite its critical function, many of the molecular mechanisms that direct the development of the ENS and the elaboration of neural network connections remain unknown. The goal of this study was to transcriptionally identify molecular pathways and candidate genes that drive specification, differentiation and the neural circuitry of specific neural progenitors, the phox2b expressing ENS cell lineage, during normal enteric nervous system development. Because ENS development is tightly linked to its environment, the transcriptional landscape of the cellular environment of the intestine was also analyzed. Thousands of zebrafish intestines were manually dissected from a transgenic line expressing green fluorescent protein under the phox2b regulatory elements [Tg(phox2b:EGFP) w37 ]. Fluorescence-activated cell sorting was used to separate GFP-positive phox2b expressing ENS progenitor and derivatives from GFP-negative intestinal cells. RNA-seq was performed to obtain accurate, reproducible transcriptional profiles and the unbiased detection of low level transcripts. Analysis revealed genes and pathways that may function in ENS cell determination, genes that may be identifiers of different ENS subtypes, and genes that define the non-neural cellular microenvironment of the ENS. Differential expression analysis between the two cell populations revealed the expected neuronal nature of the phox2b expressing lineage including the enrichment for genes required for neurogenesis and synaptogenesis, and identified many novel genes not previously associated with ENS development. Pathway analysis pointed to a high level of G-protein coupled pathway activation, and identified novel roles for candidate pathways such as the Nogo/Reticulon axon guidance pathway in ENS development. We report the comprehensive gene expression profiles of a lineage-specific population of enteric progenitors, their derivatives, and their microenvironment during normal enteric nervous system development. Our results confirm previously implicated genes and pathways required for ENS development, and also identify scores of novel candidate genes and pathways. Thus, our dataset suggests various potential mechanisms that drive ENS development facilitating characterization and discovery of novel therapeutic strategies to improve gastrointestinal disorders.


Spectroscopic and Crystallographic Evidence for the Role of a Water-Containing H-Bond Network in Oxidase Activity of an Engineered Myoglobin.
Heme-copper oxidases (HCOs) catalyze efficient reduction of oxygen to water in biological respiration. Despite progress in studying native enzymes and their models, the roles of non-covalent interactions in promoting this activity are still not well understood. Here we report EPR spectroscopic studies of cryoreduced oxy-F33Y-CuBMb, a functional model of HCOs engineered in myoglobin (Mb). We find that cryoreduction at 77 K of the O2-bound form, trapped in the conformation of the parent oxyferrous form, displays a ferric-hydroperoxo EPR signal, in contrast to the cryoreduced oxy-wild-type (WT) Mb, which is unable to deliver a proton and shows a signal from the peroxo-ferric state. Crystallography of oxy-F33Y-CuBMb reveals an extensive H-bond network involving H2O molecules, which is absent from oxy-WTMb. This H-bonding proton-delivery network is the key structural feature that transforms the reversible oxygen-binding protein, WTMb, into F33Y-CuBMb, an oxygen-activating enzyme that reduces O2 to H2O. These results provide direct evidence of the importance of H-bond networks involving H2O in conferring enzymatic activity to a designed protein. Incorporating such extended H-bond networks in designing other metalloenzymes may allow us to confer and fine-tune their enzymatic activities.


The role of CFTR channel in female infertility.
The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-activated trans-membrane ATP gated anion channel present in most epithelia, which transports chloride and bicarbonate ions across the apical membrane. Mutations in the CFTR protein are known to result in defective expression or function, notably the inhibition of chloride and bicarbonate transport. This can result in cystic fibrosis (CF), a disorder characterised by thickness of the mucus lining of the epithelial cells of the alimentary and respiratory tracts, sweat ducts and reproductive organs. As a consequence, there is a reduction in fluid transport at the apical surface. While the most devastating effect of CF is mortality, about 98% of men with CF are infertile, consequent of early blockage of or failure to develop the mesonephrotic ducts as well as the vas deferens. The effect of CF of female fertility is less well-understood. This review highlights the genetics and pathophysiology as well as the mechanism of action of CF on female infertility.


Free and acid-labile hydrogen sulfide concentrations in mouse tissues: anomalously high free hydrogen sulfide in aortic tissue.
Endogenously produced hydrogen sulfide is thought to function as an intracellular messenger. There is, however, little information on tissue concentrations of free hydrogen sulfide, the putative messenger form of this molecule, versus that of the bound (acid-labile) form. The present report describes the application of a novel technique to measure free and acid-labile hydrogen sulfide in mouse tissues. Very low free hydrogen sulfide concentrations (<0.050 μmol/kg) were observed in brain, liver, blood, heart, kidney, striated muscle, and esophagus. Aortic concentrations of free hydrogen sulfide were 20 to 100 times greater than that of the other tissues. Acid-labile hydrogen sulfide concentrations were multiple orders of magnitude greater than that of the free form in every tissue other than aorta. Previous reports of tissue hydrogen sulfide concentrations of 30 to >100 μmol/kg measured bound rather than free hydrogen sulfide, the observation that aorta contains anomalously high free hydrogen sulfide concentrations lends support for a vasodilator function for this molecule, and the very low free hydrogen sulfide concentrations in most tissues seemingly requires intermediation of a yet to be described receptor-like mechanism if this molecule is to serve as a gasotransmitter.


High frequency of colonization and absence of identifiable risk factors for methicillin-resistant Staphylococcus aureus (MRSA)in intensive care units in Brazil.
Colonization of hospitalized patients with methicillin-resistant Staphylococcus aureus (MRSA) is of increasing concern. To evaluate this problem in Intensive Care Units (ICUs) in Brazil, we studied 100 patients admitted to two ICUs from April to June, 1997. Of the 100 patients, 70 were male, 53 were age 60 years or older, 55 were previously hospitalized, 78 were transferred to the ICU from other hospital units, 49 had received antibiotic therapy, and 66 had undergone recent surgery. Nasal and axillary swab cultures were obtained on admission and every 48 hours thereafter until discharge. MRSA were identified by plating any cultured S. aureus on Mueller-Hinton agar containing 6 microg/ml of oxacillin. At the time of admission, 46 (46%) of the patients were colonized with MRSA. No associated risk factors for acquiring MRSA (age, previous hospitalization, prior surgery) could be identified. Of the 54 patients negative for MRSA on admission, 28 (52%) became colonized while in the ICU. Sixteen (22%) of the 74 colonized patients (colonized either on admission or during ICU stay) had associated respiratory or urinary tract infections due to MRSA, and 9 (56%) died. No correlation with special risk factors (invasive procedures, antibiotic use, age, chronic disease) was identified. MRSA occurred frequently, but there was minimal evidence of associated risk factors. Thus, control of MRSA cannot be accomplished by targeting special factors alone, but requires attention to preventing microbial spread in all areas. Of special concern is the high frequency of acquiring the organism in the ICU (52%). Education concerning the importance of hand washing, environmental surface cleaning, and barrier protection from infected patients is needed.


Performance of children with mental retardation after cochlear implantation: speech perception, speech intelligibility, and language development.
Children with mental retardation (MR) obtain demonstrable benefit from cochlear implantation, and their postoperative performance was tempered by the degree of MR. The purpose of this study was to investigate the performance of children with MR after implantation, and to explore their progress according to the degree of MR. Fifteen implanted children with MR were included. Progress in speech perception, speech intelligibility, and language was measured using Categories of Auditory Performance, monosyllabic word test, Speech Intelligibility Rating, and Language Scale before and after implantation. We retrospectively examined outcomes and explored the association between the progress and the degree of MR after implantation. We compared monosyllabic word test scores using repeated-measures ANOVA. Speech perception and speech intelligibility for children with mild MR improved consistently after implantation. After implantation, monosyllabic word test scores did not differ significantly between children with mild MR and children with no additional disabilities. Although language development of children with mild MR was slow, they could communicate verbally 3 years after implantation. Children with moderate MR progressed more slowly and had limitations in speech and language development, and these children could communicate by vocalization and gesture 3 years after implantation.


Epithelial-to-mesenchymal transition hinders interferon-γ-dependent immunosurveillance in lung cancer cells.
The epithelial-to-mesenchymal transition (EMT) is involved in cancer metastasis; nevertheless, interferon (IFN)-γ induces anticancer activities by causing cell growth suppression, cytotoxicity, and migration inhibition. Regarding the poor response to exogenously administered IFN-γ as anticancer therapy, it was hypothesized that malignant cells may acquire a means of escaping from IFN-γ immunosurveillance, likely through an EMT-related process. A genomic analysis of human lung cancers revealed a negative link between the EMT and IFN-γ signaling, while compared to human lung adenocarcinoma A549 cells, IFN-γ-hyporesponsive AS2 cells exhibited mesenchymal characteristics. Chemically, physically, and genetically engineered EMT attenuated IFN-γ-induced IFN regulatory factor 1 transactivation. Poststimulation of transforming growth factor-β induced the EMT and also selectively retarded IFN-γ-responsive gene expression as well as IFN-γ-induced signal transducer and activator of transcription 1 activation, major histocompatibility complex I, and CD54 expression, cell migration/invasion inhibition, and direct/indirect cytotoxicity. Without changes in IFN-γ receptors, excessive oxidative activation of Src homology-2 containing phosphatase 2 (SHP2) in cells undergoing the EMT primarily caused cellular hyporesponsiveness to IFN-γ signaling and cytotoxicity, while combining an SHP2 inhibitor or antioxidant sensitized EMT-associated AS2 and mesenchymal A549 cells to IFN-γ-induced priming effects on tumor necrosis factor-related apoptosis-inducing ligand cytotoxicity. In cell line-derived xenograft models, combined treatment with IFN-γ and an SHP2 inhibitor induced enhanced anticancer activities. These results imply that EMT-associated SHP2 activation inhibits IFN-γ signaling, facilitating lung cancer cell escape from IFN-γ immunosurveillance.


Mucopolysaccharidosis VI.
Mucopolysaccharidosis VI (MPS VI) is a lysosomal storage disease with progressive multisystem involvement, associated with a deficiency of arylsulfatase B leading to the accumulation of dermatan sulfate. Birth prevalence is between 1 in 43,261 and 1 in 1,505,160 live births. The disorder shows a wide spectrum of symptoms from slowly to rapidly progressing forms. The characteristic skeletal dysplasia includes short stature, dysostosis multiplex and degenerative joint disease. Rapidly progressing forms may have onset from birth, elevated urinary glycosaminoglycans (generally >100 microg/mg creatinine), severe dysostosis multiplex, short stature, and death before the 2nd or 3rd decades. A more slowly progressing form has been described as having later onset, mildly elevated glycosaminoglycans (generally <100 microg/mg creatinine), mild dysostosis multiplex, with death in the 4th or 5th decades. Other clinical findings may include cardiac valve disease, reduced pulmonary function, hepatosplenomegaly, sinusitis, otitis media, hearing loss, sleep apnea, corneal clouding, carpal tunnel disease, and inguinal or umbilical hernia. Although intellectual deficit is generally absent in MPS VI, central nervous system findings may include cervical cord compression caused by cervical spinal instability, meningeal thickening and/or bony stenosis, communicating hydrocephalus, optic nerve atrophy and blindness. The disorder is transmitted in an autosomal recessive manner and is caused by mutations in the ARSB gene, located in chromosome 5 (5q13-5q14). Over 130 ARSB mutations have been reported, causing absent or reduced arylsulfatase B (N-acetylgalactosamine 4-sulfatase) activity and interrupted dermatan sulfate and chondroitin sulfate degradation. Diagnosis generally requires evidence of clinical phenotype, arylsulfatase B enzyme activity <10% of the lower limit of normal in cultured fibroblasts or isolated leukocytes, and demonstration of a normal activity of a different sulfatase enzyme (to exclude multiple sulfatase deficiency). The finding of elevated urinary dermatan sulfate with the absence of heparan sulfate is supportive. In addition to multiple sulfatase deficiency, the differential diagnosis should also include other forms of MPS (MPS I, II IVA, VII), sialidosis and mucolipidosis. Before enzyme replacement therapy (ERT) with galsulfase (Naglazyme), clinical management was limited to supportive care and hematopoietic stem cell transplantation. Galsulfase is now widely available and is a specific therapy providing improved endurance with an acceptable safety profile. Prognosis is variable depending on the age of onset, rate of disease progression, age at initiation of ERT and on the quality of the medical care provided.


Independent and interactive effects of DOF affecting germination 1 (DAG1) and the Della proteins GA insensitive (GAI) and Repressor of ga1-3 (RGA) in embryo development and seed germination.
The transcription factor DOF AFFECTING GERMINATION1 (DAG1) is a repressor of seed germination acting downstream of the master repressor PHYTOCROME INTERACTING FACTOR3-LIKE 5 (PIL5). Among others, PIL5 induces the expression of the genes encoding the two DELLA proteins GA INSENSITIVE 1 (GAI) and REPRESSOR OF ga1-3 (RGA). Based on the properties of gai-t6 and rga28 mutant seeds, we show here that the absence of RGA severely increases dormancy, while lack of GAI only partially compensates RGA inactivation. In addition, the germination properties of the dag1rga28 double mutant are different from those of the dag1 and rga28 single mutants, suggesting that RGA and DAG1 act in independent branches of the PIL5-controlled germination pathway. Surprisingly, the dag1gai-t6 double mutant proved embryo-lethal, suggesting an unexpected involvement of (a possible complex between) DAG1 and GAI in embryo development. Rather than overlapping functions as previously suggested, we show that RGA and GAI play distinct roles in seed germination, and that GAI interacts with DAG1 in embryo development.


Role of cranberry juice on molecular-scale surface characteristics and adhesion behavior of Escherichia coli.
Cranberry juice has long been believed to benefit the prevention and treatment of urinary tract infections (UTIs). As the first step in the development of infection, bacterial adhesion is of great research interest, yet few studies have addressed molecular level adhesion in this context. P-fimbriated Escherichia coli play a major role in the development of a serious type of UTI, acute pyelonephritis. Experiments were conducted to investigate the molecular-scale effects of cranberry juice on two E. coli strains: HB101, which has no fimbriae, and the mutant HB101pDC1 which expresses P-fimbriae. Atomic force microscopy (AFM) was used to investigate both bacterial surface characteristics and adhesion forces between a probe surface (silicon nitride) and the bacteria, providing a direct evaluation of bacterial adhesion and interaction forces. Cranberry juice affected bacterial surface polymer and adhesion behavior after a short exposure period (<3 h). Cranberry juice affected the P-fimbriated bacteria by decreasing the adhesion forces between the bacterium and tip and by altering the conformation of the surface macromolecules on E. coli HB101pDC1. The equilibrium length of polymer (P-fimbriae) on this bacterium decreased from approximately 148 to approximately 48 nm upon being exposed to cranberry juice. Highly acidic conditions were not necessary for the prevention of bacterial adhesion, since neutralization of cranberry juice solutions to pH = 7.0 allowed us to observe differences in adhesion between the E. coli strains. Our results demonstrate molecular-level changes in the surfaces of P-fimbriated E. coli upon exposure to neutralized cranberry juice.


Purification and characterization of bovine aorta ribonucleases.
1. Two ribonucleases (aorta ribonuclease I and aorta ribonuclease II) from bovine aorta were purified 4611-fold and 667-fold respectively. Ethanolic precipitation, acid extraction, isoionic precipitation at pH3.5 and Bio-Rex 70 column chromatography were the methods employed. 2. Aorta ribonuclease I exhibited no deoxyribonuclease or alkaline phosphatase activity. 3. Aorta ribonuclease I appeared to be homogeneous when subjected to discontinuous gel electrophoresis. 4. Aorta ribonuclease II exhibited the same properties as aorta ribonuclease previously isolated. 5. The activities of the aorta ribonucleases and pancreatic ribonuclease on homopolymers and dinucleoside phosphates were compared. 6. Aorta ribonuclease I exhibited optimum pH7.5 and, under the assay conditions used, optimum temperature 60 degrees .


p53 nuclear accumulation in preneoplastic lesions and stage I uterine endometrioid adenocarcinoma.
A total of 118 endometrial neoplastic and preneoplastic lesions comprising 43 uterine endometrioid adenocarcinoma at stage I, 40 complex (adenomatous) hyperplasias and 35 atypical hyperplasias were examined for p53 nuclear accumulation to assess the incidence of p53 alterations in infiltrating carcinomas and to verify if p53 aberrations may allow the identification of a subset of premalignant cases with high risk of progression. No specific immunostaining was observed in the cases of complex hyperplasia without atypias. One (3%) of 35 atypical hyperplasias showed focal areas of p53 immuno-reactivity. The overall frequency of p53 overexpression in endometrial carcinomas was 54%. The distribution of cases with nuclear accumulation of p53 was significantly different (p=0.01) in tumours with different degree of invasiveness. In addition, p53 nuclear accumulation was observed more often in tumours with moderate (G2) or poor differentiation (G3) (p=0.03). Our data indicate that p53 aberrations are not early events in endometrial carcinogenesis and may be related with tumour progression and aggressiveness.


GWAS meta-analysis followed by Mendelian randomization revealed potential control mechanisms for circulating α-Klotho levels.
The protein α-Klotho acts as transmembrane co-receptor for fibroblast growth factor 23 (FGF23) and is a key regulator of phosphate homeostasis. However, α-Klotho also exists in a circulating form, with pleiotropic, but incompletely understood functions and regulation. Therefore, we undertook a genome-wide association study (GWAS) meta-analysis followed by Mendelian randomization (MR) of circulating α-Klotho levels. Plasma α-Klotho levels were measured by enzyme-linked immunosorbent assay (ELISA) in the Ludwigshafen Risk and Cardiovascular Health and Avon Longitudinal Study of Parents and Children (mothers) cohorts, followed by a GWAS meta-analysis in 4376 individuals across the two cohorts. Six signals at five loci were associated with circulating α-Klotho levels at genome-wide significance (P < 5 × 10-8), namely ABO, KL, FGFR1, and two post-translational modification genes, B4GALNT3 and CHST9. Together, these loci explained >9% of the variation in circulating α-Klotho levels. MR analyses revealed no causal relationships between α-Klotho and renal function, FGF23-dependent factors such as vitamin D and phosphate levels, or bone mineral density. The screening for genetic correlations with other phenotypes followed by targeted MR suggested causal effects of liability of Crohn's disease risk [Inverse variance weighted (IVW) beta = 0.059 (95% confidence interval 0.026, 0.093)] and low-density lipoprotein cholesterol levels [-0.198 (-0.332, -0.063)] on α-Klotho. Our GWAS findings suggest that two enzymes involved in post-translational modification, B4GALNT3 and CHST9, contribute to genetic influences on α-Klotho levels, presumably by affecting protein turnover and stability. Subsequent evidence from MR analyses on α-Klotho levels suggest regulation by mechanisms besides phosphate-homeostasis and raise the possibility of cross-talk with FGF19- and FGF21-dependent pathways, respectively. Significance statement: α-Klotho as a transmembrane protein is well investigated along the endocrine FGF23-α-Klotho pathway. However, the role of the circulating form of α-Klotho, which is generated by cleavage of transmembrane α-Klotho, remains incompletely understood. Genetic analyses might help to elucidate novel regulatory and functional mechanisms. The identification of genetic factors related to circulating α-Klotho further enables MR to examine causal relationships with other factors. The findings from the first GWAS meta-analysis of circulating α-Klotho levels identified six genome-wide significant signals across five genes. Given the function of two of the genes identified, B4GALNT3 and CHST9, it is tempting to speculate that post-translational modification significantly contributes to genetic influences on α-Klotho levels, presumably by affecting protein turnover and stability.


Mammalian target of rapamycin (mTOR): a central regulator of male fertility?
Mammalian target of rapamycin (mTOR) is a central regulator of cellular metabolic phenotype and is involved in virtually all aspects of cellular function. It integrates not only nutrient and energy-sensing pathways but also actin cytoskeleton organization, in response to environmental cues including growth factors and cellular energy levels. These events are pivotal for spermatogenesis and determine the reproductive potential of males. Yet, the molecular mechanisms by which mTOR signaling acts in male reproductive system remain a matter of debate. Here, we review the current knowledge on physiological and molecular events mediated by mTOR in testis and testicular cells. In recent years, mTOR inhibition has been explored as a prime strategy to develop novel therapeutic approaches to treat cancer, cardiovascular disease, autoimmunity, and metabolic disorders. However, the physiological consequences of mTOR dysregulation and inhibition to male reproductive potential are still not fully understood. Compelling evidence suggests that mTOR is an arising regulator of male fertility and better understanding of this atypical protein kinase coordinated action in testis will provide insightful information concerning its biological significance in other tissues/organs. We also discuss why a new generation of mTOR inhibitors aiming to be used in clinical practice may also need to include an integrative view on the effects in male reproductive system.


Regulation of corneal inflammation by neutrophil-dependent cleavage of keratan sulfate proteoglycans as a model for breakdown of the chemokine gradient.
Keratocan and lumican are small, leucine-rich repeat KSPGs in the extracellular matrix (ECM) of the mammalian cornea, whose primary role is to maintain corneal transparency. In the current study, we examined the role of these proteoglycans in the breakdown of the chemokine gradient and resolution of corneal inflammation. LPS was injected into the corneal stroma of C57BL/6 mice, and corneal extracts were examined by immunoblot analysis. We found reduced expression of the 52-kD keratocan protein after 6 h and conversely, increased expression of 34/37 kD immunoreactive products. Further, appearance of the 34/37-kD proteins was dependent on neutrophil infiltration to the cornea, as the appearance of these products was coincident with neutrophil infiltration, and the 34/37-kD products were not detected in explanted corneas or in CXCR2(-/-) corneas with deficient neutrophil recruitment. Furthermore, the 34/37-kD products and CXCL1/KC were detected in the anterior chamber, into which the corneal stroma drains; and CXCL1/KC was elevated significantly in keratocan(-/-) and lumican(-/-) mice. Together, these findings indicate that the inflammatory response in the cornea is regulated by proteoglycan/CXCL1 complexes, and their diffusion into the anterior chamber is consistent with release of a chemokine gradient and resolution of inflammation.


Magnetic resonance spectroscopy in affective disorders.
Magnetic resonance spectroscopy (MRS) is a non-invasive tool for in vivo chemical analysis that has been applied to neurobiological or psychopharmacological studies of affective disorders. Studies employing 31P-MRS and 1H-MRS have indicated possible abnormalities in membrane phospholipid metabolism, high-energy phosphate metabolism, and intracellular pH in affective disorders. They have also suggested that lithium increases the phosphomonoester (possibly inositol-1-phosphate) peak in the brain but does not increase that of choline-containing compounds in the brain. Studies employing 7Li-MRS and 19F-MRS have elucidated the pharmacokinetic properties of lithium, fluoxetine, and fluvoxamine in the brain in patients treated with these drugs.


Activating the Antitumor Immune Response in Non-Hodgkin Lymphoma Using Immune Checkpoint Inhibitors.
Non-Hodgkin lymphomas comprise a heterogenous group of disorders which differ in biology. Although response rates are high in some groups, relapsed disease can be difficult to treat, and newer approaches are needed for this patient population. It is increasingly apparent that the immune system plays a significant role in the propagation and survival of malignant cells. Immune checkpoint blocking agents augment cytotoxic activity of the adaptive and innate immune systems and enhance tumor cell killing. Anti-PD-1 and anti-CTLA-4 antibodies have been tested as both single agents and combination therapy. Although success rates with anti-PD-1 antibodies are high in patients with Hodgkin lymphoma, the results are yet to be replicated in those with non-Hodgkin lymphomas. Some lymphoma histologies, such as primary mediastinal B cell lymphoma (PMBL), central nervous system, and testicular lymphomas and gray zone lymphoma, respond favorably to PD-1 blockade, but the response rates in most lymphoma subtypes are low. Other agents including those targeting the adaptive immune system such as TIM-3, TIGIT, and BTLA and innate immune system such as CD47 and KIR are therefore in trials to test alternative ways to activate the immune system. Patient selection based on tumor biology is likely to be a determining factor in treatment response in patients, and further research exploring optimal patient populations, newer targets, and combination therapy as well as identifying biomarkers is needed.


Myxovirus-like structures in a case of human chronic polymyositis.
Intranuclear and intracytoplasmic aggregates of filaments with tubular structures and transverse striations occurred in muscle tissues biopsied from a patient with chronic polymyositis. The filamentous tubules bear a close resemblance to the incomplete form of myxovirus in which the envelope is missing. Three biopsies from the same patient, taken during a period of 1(1/2) years, all revealed these structures. This finding provides presumptive evidence that a chronic persistent viral infection may be involved in the pathogenesis of chronic polymyositis.


New techniques on deformed image motion estimation and compensation.
In this paper, new techniques for deformed image motion estimation and compensation using variable-size block-matching are proposed, which can be applied to an image sequence compression system or a moving object recognition system. The motion estimation and compensation techniques have been successfully applied in the area of image sequence coding. Many research papers on improving the performance of these techniques have been published; many directions are proposed, which can all lead to better performance than the conventional techniques. Among them, both generalized block-matching and variable-size block-matching are successfully applied in reducing the data rate of compensation error and motion information, respectively. These two algorithms have their merits, but suffer from their drawbacks. Moreover, reducing the data rate in compensation error is sometimes increasing the data rate in motion information, or vice versa. Based on these two algorithms, we propose and examine several algorithms which are effective in reducing the data rate. We then incorporate these algorithms into a system, in which they work together to overcome the disadvantages to individual and keep their merits at the same time. The proposed system can optimally balance the amount of data rate in two aspects (i.e., compensation error and motion information). Experimental results show that the proposed system outweighs the conventional techniques. Since we propose a recovery operation which tries to recover the incorrect motion vectors from the global motion, this proposed system can also be applied to the moving object recognition in image sequences.


Degree of vertical integration between the undergraduate program and clinical internship with respect to lumbopelvic diagnostic and therapeutic procedures taught at the canadian memorial chiropractic college.
The objective of this study was to determine to what extent the diagnostic and therapeutic procedures taught in the undergraduate program used for patients with lumbopelvic conditions are expected to be utilized by students during their clinical internship program at Canadian Memorial Chiropractic College or are being used by the clinical faculty. A confidential survey was distributed to clinical faculty at the college. It consisted of a list of diagnostic and therapeutic procedures used for lumbopelvic conditions taught at that college. Clinicians were asked to indicate the frequency with which they performed or they required students to perform each item. Seventeen of 23 clinicians responded. The following procedures were most likely required to be performed by clinicians: posture; ranges of motion; lower limb sensory, motor, and reflex testing; and core orthopedic tests. The following were less likely to be required to be performed: Waddell testing, Schober's test, Gillet tests, and abdominal palpation. Students were expected to perform (or clinicians performed) most of the mobilization (in particular, iliocostal, iliotransverse, and iliofemoral) and spinal manipulative therapies (in particular, the procedures referred to as the lumbar roll, lumbar pull/hook, and upper sacroiliac) taught at the college. This study suggests that there was considerable, but not complete, vertical integration between the undergraduate and clinical education program at this college.


Next Generation Sequencing and mass spectrometry reveal high taxonomic diversity and complex phytoplankton-phycotoxins patterns in Southeastern Pacific fjords.
In fjord systems, Harmful Algal Blooms (HABs) not only constitute a serious problem when affecting the wildlife and ecosystems, but also human health and economic activities related to the marine environment. This is mostly due to a broad spectrum of toxic compounds produced by several members of the phytoplankton. Nevertheless, a deep coverage of the taxonomic diversity and composition of phytoplankton species and phycotoxin profiles in HAB prone areas are still lacking and little is known about the relationship between these fundamental elements for fjord ecosystems. In this study, a detailed molecular and microscopic characterization of plankton communities was performed, together with an analysis of the occurrence and spatial patterns of lipophilic toxins in a HAB prone area, located in the Southeastern Pacific fjord region. Microscopy and molecular analyses based on the 18S rRNA gene fragment indicated high diversity and taxonomic homogeneity among stations. Four toxigenic genera were identified: Pseudo-nitzschia, Dinophysis, Prorocentrum, and Alexandrium. In agreement with the detected species, liquid chromatography coupled with mass spectrometry revealed the presence of domoic acid (DA), pectenotoxin-2 (PTX-2), dinophysistoxin-2 (DTX-2), and 13-desmethyl spirolide C (SPX-1). Furthermore, a patchy distribution among DA in different net haul size fractions was found. Our results displayed a complex phytoplankton-phycotoxin pattern and for the first time contribute to the characterization of high-resolution phytoplankton community composition and phycotoxin distribution in fjords of the Southeastern Pacific region.


MRI of stroke using hyperpolarized 129Xe.
Because there is no background signal from xenon in biological tissue, and because inhaled xenon is delivered to the brain by blood flow, we would expect a perfusion deficit, such as is seen in stroke, to reduce the xenon concentration in the region of the deficit. Thermal polarization yields negligible xenon signal relative to hyperpolarized xenon; therefore, hyperpolarized xenon can be used as a tracer of cerebral blood flow. Using a rat permanent right middle cerebral artery occlusion model, we demonstrated that hyperpolarized (129)Xe MRI is able to detect, in vivo, the hypoperfused area of focal cerebral ischemia, that is the ischemic core area of stroke. To the best of our knowledge, this is the first time that hyperpolarized (129)Xe MRI has been used to explore normal and abnormal cerebral perfusion. Our study shows a novel application of hyperpolarized (129)Xe MRI for imaging stroke, and further demonstrates its capacity to serve as a complementary tool to proton MRI for the study of the pathophysiology during brain hypoperfusion.


TMEM100 Regulates Neuropathic Pain by Reducing the Expression of Inflammatory Factors.
There is no effective treatment for peripheral nerve injury-induced chronic neuropathic pain (NP), which profoundly impacts the quality of life of those affected. Transmembraneprotein100 (TMEM100) is considered to be a pain regulatory protein and is expressed in the dorsal root ganglion (DRG) of rats. However, the mechanism of pain regulation and the expression of TMEM100 following various peripheral nerve injuries are unclear. In this study, we constructed two pain models of peripheral nerve injury: tibial nerve injury (TNI) and chronic constriction injury (CCI). This study found that the Paw Withdrawal Mechanical Threshold (PWMT) and Paw Withdraw Thermal Latency (PWTL) of the rats in the two pain models decreased significantly, and the expression of TMEM100 in the DRG of two groups also decreased significantly. Furthermore, the decrease in the CCI group was more obvious than in the TNI group. There was no significant statistical significance (P > 0.05). We constructed an adeno-associated virus 6 (AAV6) vector expressing recombinant fluorescent TMEM100 protein and injected it into the sciatic nerve (SN) of two pain models: CCI and TNI. PWMT and PWTL were significantly increased in the two groups, along with the expression of TMEM100 in the spinal cord and DRG. It also significantly inhibited the activation of microglia, astrocytes, and several inflammatory mediators (TNF- α, IL-1 β, and IL-6). In summary, the results of this study suggested that TMEM100 might be a promising molecular strategy for the treatment of NP, and its anti-inflammatory effects might play an important role in pain relief.


Late Effects of Clubfoot Deformity in Adolescent and Young Adult Patients Whose Initial Treatment Was an Extensive Soft-tissue Release: Topic Review and Clinical Case Series.
Children with congenital clubfoot often have residual deformity, pain, and limited function in adolescence and young adulthood. These patients represent a heterogeneous group that often requires an individualized management strategy. This article reviews the available literature on this topic while proposing a descriptive classification system based on a review of patients at our institution who underwent surgery for problems related to previous clubfoot deformity during the period between January 1999 and January 2012. Seventy-two patients (93 feet) underwent surgical treatment for the late effects of clubfoot deformity at an average age of 13 years (range 9 to 19 years). All patients had been treated at a young age with serial casting, and most had at least one previous surgery on the affected foot or feet. Five common patterns of pathology identified were as follows: undercorrection, overcorrection, dorsal bunion, anterior ankle impingement, and lateral hindfoot impingement. Management pathways for each group of the presenting problems is described. To our knowledge, this topic review represents the largest report of adolescent and young adult patients with residual clubfoot deformity in the literature.


[The dental filling therapy of carious deciduous molars: materials and methods. A review].
There is little consensus as to which restorative material or techniques are best suited to restore primary teeth. Techniques developed for the permanent dentition can seldom be applied to the primary teeth because of morphological differences and lack of cooperation by the children. The morphological properties of primary molars are discussed in this paper in relation to their therapeutic consequences. In addition to general guidelines for dental therapy in children, current restorative materials and techniques are compared regarding their suitability for the primary dentition. Finally the goals for the development of an ideal restorative material are formulated.


Prevalence of Misinformation and Factchecks on the COVID-19 Pandemic in 35 Countries: Observational Infodemiology Study.
The COVID-19 pandemic has been accompanied by an infodemic, in which a plethora of false information has been rapidly disseminated online, leading to serious harm worldwide. This study aims to analyze the prevalence of common misinformation related to the COVID-19 pandemic. We conducted an online survey via social media platforms and a survey company to determine whether respondents have been exposed to a broad set of false claims and fact-checked information on the disease. We obtained more than 41,000 responses from 1257 participants in 85 countries, but for our analysis, we only included responses from 35 countries that had at least 15 respondents. We identified a strong negative correlation between a country's Gross Domestic Product per-capita and the prevalence of misinformation, with poorer countries having a higher prevalence of misinformation (Spearman ρ=-0.72; P<.001). We also found that fact checks spread to a lesser degree than their respective false claims, following a sublinear trend (β=.64). Our results imply that the potential harm of misinformation could be more substantial for low-income countries than high-income countries. Countries with poor infrastructures might have to combat not only the spreading pandemic but also the COVID-19 infodemic, which can derail efforts in saving lives.


Increase in incidence of hyperthyroidism predominantly occurs in young people after iodine fortification of salt in Denmark.
To prevent goiter and nodular hyperthyroidism, iodine fortification (IF) of salt was introduced in Denmark in 1998. We prospectively registered all new cases of overt hyperthyroidism in two areas of Denmark before and for the first 6 yr after iodine fortification. We used a computer-based register of all new cases of hyperthyroidism in two population subcohorts with moderate iodine deficiency (Aalborg, n = 310,124) and mild iodine deficiency (Copenhagen, n = 225,707), respectively. Data were obtained 1) before IF (1997-1998); 2) during voluntary IF (1999-2000); 3) during the early (2001-2002) period of mandatory IF; and 4) during the late (2003-2004) period with mandatory IF. The overall incidence rate of hyperthyroidism increased [baseline, 102.8/100,000/year; voluntary IF, 122.8; early mandatory IF, 140.7; late mandatory IF, 138.7 (P for trend <0.001)]. Hyperthyroidism increased in both sexes (P < 0.001) and in all age groups: 0-19, 20-39, 40-59, and 60+ yr (P for trend <0.001). The increase was relatively highest in young adults aged 20-39 yr: late mandatory IF (percent increase from baseline), age 20-39, 160%, P < 0.001; age 40-59, 29%, P < 0.01; age 60+ yr, 13%, P = not significant. Even a cautious iodization of salt results in an increase in the incidence rate of hyperthyroidism. Contrary to current concepts, many of the new cases were observed in young subjects, and are presumably of autoimmune origin. Furthermore, monitoring is expected to show a decrease in the number of elderly subjects suffering from nodular hyperthyroidism.


Long-term increases in tropical flowering activity across growth forms in response to rising CO2 and climate change.
Mounting evidence suggests that anthropogenic global change is altering plant species composition in tropical forests. Fewer studies, however, have focused on long-term trends in reproductive activity, in part because of the lack of data from tropical sites. Here, we analyze a 28-year record of tropical flower phenology in response to anthropogenic climate and atmospheric change. We show that a multidecadal increase in flower activity is most strongly associated with rising atmospheric CO2 concentrations using yearly aggregated data. Compared to significant climatic factors, CO2 had on average an approximately three-, four-, or fivefold stronger effect than rainfall, solar radiation, and the Multivariate ENSO Index, respectively. Peaks in flower activity were associated with greater solar radiation and lower rainfall during El Niño years. The effect of atmospheric CO2 on flowering has diminished over the most recent decade for lianas and canopy trees, whereas flowering of midstory trees and shrub species continued to increase with rising CO2 . Increases in flowering were accompanied by a lengthening of flowering duration for canopy and midstory trees. Understory treelets did not show increases in flowering but did show increases in duration. Given that atmospheric CO2 will likely continue to climb over the next century, a long-term increase in flowering activity may persist in some growth forms until checked by nutrient limitation or by climate change through rising temperatures, increasing drought frequency and/or increasing cloudiness and reduced insolation.