Relationship between pharyngeal response time and lateralized brain lesion in stroke.
Objectives: To analyze the relationship between pharyngeal response time (PRT) and lateralization of brain lesions. Methods: A Cross-sectional study. 73 videofluoroscopic swallow studies (VFSS)were conducted on patients after stroke The study subjects were divided into : group 1 (G1) consisting of 39 individuals with left cortical lesion and group 2 (G2) consisting of 34 individuals with right cortical lesions. The VFSS of G1 and G2 subjects were analyzed using puree (A) and liquid (B) consistencies, and were also subdivided into young adults and older persons. . The mean PRT was divided into times shorter and longer than 250 ms. Results: No statistically significant difference was observed between G1 and G2 for the A and B consistencies, being obtained : G1 (A mean: 56.6 ms; B mean: 99.5 ms; A mean: 3627 ms; B mean: 1712 ms) and G2 (A mean: 79.6 ms; B mean: 110.7 ms; A mean: 2040 ms, B mean: 1529 ms), for PRT shorter (A:p = .673; B: p = 1.000) and longer (A: p = .435; B: p = .847) than 250 ms, respectively. No statistically significant difference was found regarding the variable age in the comparison between young and old adults for mean PRT according to the A and B consistencies in G1 (A: p = .260; B: p = .732) and G2 (A: p = .586; B: p = .104). Conclusion: No relationship between PRT and lateralization of brain lesion was observed with respect to subject age and to the swallowing of different food consistencies.


Clearing the smoke: are tobacco cessation interventions by the dental team successful?
This study aimed to assess the outcomes of tailored tobacco cessation interventions in a dental clinic setting over one year. This interventional study recruited 1206 tobacco users who attended a smoking cessation clinic within an Oral Medicine Department in Bangalore, India. Baseline characteristics were recorded followed by a tailored smoking cessation intervention, delivered by trained faculty members and students. This followed the 3Es and 6As model - including a combination of personalised counselling, pharmacotherapy and one year follow up. 1206 participants were recruited. 95% were already receiving treatment within the dental hospital and 5% were walk-in patients, attempting to quit tobacco use. Participants who had been smoking or using smokeless tobacco more than twice daily for greater than 6 months were included in the study. Participants were excluded from the study if they were currently using nicotine replacement therapy (NRT) or non-NRT therapy for tobacco cessation, pregnant and lactating women, a recent history of myocardial infarction or a history of gastric ulcers. Outcomes were categorised into the following four groups; Complete abstinence, Harm reduction (>50% reduction), No change and lost to follow up. Of the patients who completed the 12 month follow up, 18% quit tobacco use (n = 180), 34.2% reduced their tobacco usage by greater than 50% (n = 342), 41.5% exhibited no change (n = 415) and 6.2% relapsed (n = 62). Salivary nicotine levels were used to confirm self-reported abstinence. This study shows that dental professional led tobacco cessation programmes can be successful in supporting patients to quit or reduce tobacco usage.


Clinico-Etiological Evaluation of Chronic Leg Ulcer in a Tertiary Care Center of Eastern India.
A chronic leg ulcer (CLU) is a significant public health problem. It has various etiologies. Racial, familial, occupational, and social factors may also have an impact on the prevalence of different causes of leg ulcers. Though there are western data on the epidemiology of leg ulcer, similar data are largely unavailable from our part of the world. We undertook a study in a tertiary care center in eastern India to determine the clinical and etiological pattern of patients with CLU. Hundred consecutive patients presenting with CLU, fulfilling the criteria, were included after informed consent. Patients were subjected to proper history taking, clinical examination, routine blood test, and pus for culture and sensitivity test (where needed) along with Ankle Brachial Index (ABI). Among the 100 patients, venous ulcer (34%) was predominant followed by arterial ulcer (14%), mixed arterial and venous ulcer (11%). History of smoking (56%) and obesity (BMI >25) (32%) were the common risk factors in leg ulcer patients. Fifty nine percent of the total CLU were infected and out of this, 86.4% showed growth of microorganisms. Staphylococcus aureus (39%) was the most commonly isolated organism, followed by Pseudomonas aeruginosa (15%). Eleven (24.44%) clinically diagnosed venous ulcer patients showed significantly lower ABI (<0.9) and were diagnosed as mixed ulcer (a venous ulcer with a peripheral arterial disease). Venous ulcer and mixed ulcer are the most common type of CLU.


Schistosoma mansoni: assessment of effects of oleic acid, cercarial age and water temperature on parasite-host attraction.
Although the lifecycle of Schistosoma spp. and pathophysiology of schistosomiasis have been established, the mechanism by which cercariae find their host is not well understood. Speculatively, host infection by random and accidental host contact is not as biologically plausible as a biochemical mechanism of mammalian attraction. A few studies have indicated that biochemical cues and temperature gradients may play a role in host identification, attraction and attachment triggers. This study aimed to elucidate these mechanisms more specifically through evaluation of biochemical, age and temperature influences leading to Schistosoma mansoni cercariae attraction and attachment behaviors. Oleic acid, a common unsaturated free fatty acid in the outer layer of human skin, was tested for cercariae attraction across biologically relevant concentrations. Influence of media type (beeswax, nail varnish and agar), age-dependent behavior variability and environmentally appropriate temperatures (22 and 30 °C) were also evaluated. Results indicated that oleic acid at concentrations of 0.3, 0.9 and 1.8 g/mL in beeswax significantly increased median attachment to media (median attachment of 7.50%, 4.20% and 3.71%, respectively, P<0.001), compared with plain beeswax, with maximal attachment of 30.30% at 0.3g/mL of oleic acid. In media containing 0.3 g/mL of oleic acid, cercarial attachment was highest for freshly emerged cercariae to 5h post-emergence, with a significant decrease in attachment behavior at 10h post-emergence (P<0.01). Aquatic temperature at which cercariae were exposed to media did not yield significant results (P value >0.05). Biochemical, age and environmental factors influencing cercarial host attraction and attachment behavior have been elucidated by this study. This information will inform further development of devices for environmental surveillance and potentially improve cercarial exposure prevention strategies.


Structural properties of thin-film ferromagnetic topological insulators.
We present a comprehensive study of the crystal structure of the thin-film, ferromagnetic topological insulator (Bi, Sb)2-x V x Te3. The dissipationless quantum anomalous Hall edge states it manifests are of particular interest for spintronics, as a natural spin filter or pure spin source, and as qubits for topological quantum computing. For ranges typically used in experiments, we investigate the effect of doping, substrate choice and film thickness on the (Bi, Sb)2Te3 unit cell using high-resolution X-ray diffractometry. Scanning transmission electron microscopy and energy-dispersive X-ray spectroscopy measurements provide local structural and interfacial information. We find that the unit cell is unaffected in-plane by vanadium doping changes, and remains unchanged over a thickness range of 4-10 quintuple layers (1 QL ≈ 1 nm). The in-plane lattice parameter (a) also remains the same in films grown on different substrate materials. However, out-of-plane the c-axis increases with the doping level and thicknesses >10 QL, and is potentially reduced in films grown on Si (1 1 1).


[Establishment of Patient-Derived Xenograft (PDX) Zebrafish Model of Multiple Myeloma and Its Application in Drug Screening].
To establish a MM patient-derived tumor xenograft model (MM-PDX) in zebrafish, and to evaluate the anti-myeloma activity of indirubin-3'-monoxime(I3MO) using this model. Zebrafish embryos 2 days after fertilization were transplanted with fluorescence labeled myeloma primary tumor cells, the survival of primary tumor cells in zebrafish was observed at 0,16 and 24 hours after cell injection. The zebrafish embryos after tumor cell transplantation were randomly divided into control group, BTZ treatment and I3MO treatment group. Before and 24 hours after treatment with BTZ and I3MO, the positive area with calcein or Dil in zebrafish were observed under fluorescence microscope to reflect the survival of tumor cells, and it was verified. MM patient derived tumor cells survived in zebrafish. The construction of MM-PDX was successful. Compared with control group, the fluo- rescence area of the BTZ and I3MO treatment groups in zebrafish were significantly decreased(P<0.05), and BTZ and I3MO significantly inhibited the survival of MM cells in zebrafish. MM-PDX model was successfully established. Zebrafish model derived from tumor cells of MM patients can be used as a tool for drug screening of MM.


The role of polo-like kinase 1 in carcinogenesis: cause or consequence?
Polo-like kinase 1 (Plk1) is a well-established mitotic regulator with a diverse range of biologic functions continually being identified throughout the cell cycle. Preclinical evidence suggests that the molecular targeting of Plk1 could be an effective therapeutic strategy in a wide range of cancers; however, that success has yet to be translated to the clinical level. The lack of clinical success has raised the question of whether there is a true oncogenic addiction to Plk1 or if its overexpression in tumors is solely an artifact of increased cellular proliferation. In this review, we address the role of Plk1 in carcinogenesis by discussing the cell cycle and DNA damage response with respect to their associations with classic oncogenic and tumor suppressor pathways that contribute to the transcriptional regulation of Plk1. A thorough examination of the available literature suggests that Plk1 activity can be dysregulated through key transformative pathways, including both p53 and pRb. On the basis of the available literature, it may be somewhat premature to draw a definitive conclusion on the role of Plk1 in carcinogenesis. However, evidence supports the notion that oncogene dependence on Plk1 is not a late occurrence in carcinogenesis and it is likely that Plk1 plays an active role in carcinogenic transformation.


Early mortality in German patients with lung cancer: risk factors associated with 30-and 60-day mortality.
Despite therapeutic advances, early mortality in lung cancer is still prevalent. In this study, we aimed to assess risk factors for 30- and 60-day mortality in German lung cancer patients. In this retrospective cross-sectional analysis, we used data of lung cancer patients treated at LMU Hospital Munich between 2015 and 2019. We categorized patients into 30-day mortality, 60 day-mortality, and longer survival. We used Student's t-test and ANOVA to compare means and Chi2-test to compare frequencies. We used logistic regression analysis to identify factors associated with a risk for early mortality. Of the 2454 lung cancer patients, 2.0% (n = 50) died within 30 and 1.7% (n = 41) within 30 to 60 days of diagnosis. Older age and advanced stage at diagnosis were significantly associated with early mortality in the univariate and the multivariate analysis. Patients in the 30-day mortality group significantly more often did not receive tumor-directed therapy. They were also more likely to die in an acute care setting compared to the 60-day mortality group. The group of patients who died unexpectedly (12.0%) was dominantly female, with a high proportion of patients with unintentional weight loss at the time of diagnosis. Our results suggest that in the treatment of patients with lung cancer there is a need for a greater focus on older patients. Moreover, physicians should pay special attention to females with recent weight loss and patients with a comorbidity of diabetes mellitus or renal impairment. Engaging a case manager focused on detecting patients with the above characteristics could help improve overall care.


Assessment of the spatial pattern of colorectal tumour perfusion estimated at perfusion CT using two-dimensional fractal analysis.
The aim was to evaluate the feasibility of fractal analysis for assessing the spatial pattern of colorectal tumour perfusion at dynamic contrast-enhanced CT (perfusion CT). Twenty patients with colorectal adenocarcinoma underwent a 65-s perfusion CT study from which a perfusion parametric map was generated using validated commercial software. The tumour was identified by an experienced radiologist, segmented via thresholding and fractal analysis applied using in-house software: fractal dimension, abundance and lacunarity were assessed for the entire outlined tumour and for selected representative areas within the tumour of low and high perfusion. Comparison was made with ten patients with normal colons, processed in a similar manner, using two-way mixed analysis of variance with statistical significance at the 5% level. Fractal values were higher in cancer than normal colon (p < or = 0.001): mean (SD) 1.71 (0.07) versus 1.61 (0.07) for fractal dimension and 7.82 (0.62) and 6.89 (0.47) for fractal abundance. Fractal values were lower in 'high' than 'low' perfusion areas. Lacunarity curves were shifted to the right for cancer compared with normal colon. In conclusion, colorectal cancer mapped by perfusion CT demonstrates fractal properties. Fractal analysis is feasible, potentially providing a quantitative measure of the spatial pattern of tumour perfusion.


Intact blood-perilymph barrier in the rat after impulse noise trauma.
The permeability of the blood-labyrinth barrier for radioactive mannitol was unchanged after impulse noise trauma. The present findings are contradictory to the theory of an increased permeability in the blood-labyrinth barrier as a result of extensive noise exposure. Noise trauma is reported to cause multiple effects on the cochlea including mechanical and metabolic damage. The aim of the study was to observe the effects of impulse noise on cochlear homeostasis. A well-established rat model was used for evaluation of the early effects of impulse noise trauma on the integrity of the blood-perilymph barrier. To evaluate whether a blood-perilymph barrier disruption contributes to cochlear injury after impulse noise, the paracellular transport of radioactive mannitol into scala vestibuli perilymph (PLV) and electrolyte concentration in perilymph were estimated. Thirteen animals exposed to synthesized impulses of 160 dB SPL peak value, at a rate of 100 pulses, were designed as the study group and 15 rats not exposed to noise were designed as the control group. After mannitol infusion each ear of the animal in the study group was separately exposed to impulse noise and PLV samples were taken during 2 h post-infusion. In the control group, corresponding PLV samples were taken after mannitol injection. At 2 h after mannitol infusion there was no difference in PLV mannitol concentration in the study group and control group (21.5%+/-2.2 and 20.5%+/-2.1, respectively). Impulse noise had no effect on the electrochemical composition of PLV.


Informed Consent for Web Paradata Use.
Survey researchers are making increasing use of paradata - such as keystrokes, clicks, and timestamps - to evaluate and improve survey instruments but also to understand respondents and how they answer surveys. Since the introduction of paradata, researchers have been asking whether and how respondents should be informed about the capture and use of their paradata while completing a survey. In a series of three vignette-based experiments, we examine alternative ways of informing respondents about capture of paradata and seeking consent for their use. In all three experiments, any mention of paradata lowers stated willingness to participate in the hypothetical surveys. Even the condition where respondents were asked to consent to the use of paradata at the end of an actual survey resulted in a significant proportion declining. Our research shows that requiring such explicit consent may reduce survey participation without adequately informing survey respondents about what paradata are and why they are being used.


[Influence of GABAergic neurons in the brain on central effects of kainate on the cardiovascular system].
Microinjection of kainate (KA 0.125, 0.25, or 0.5 microgram), a special agonist for KA receptor, into rat lateral cerebral ventricle produced dose-dependent increases in blood pressure and heart rate. Pretreatment with semicarbazide (SCZ 140 mg/kg, ip), an inhibitor of GABA biosynthesis, or picrotoxin (PIC 1 mg/kg, iv), an antagonist for GABA, significantly enhanced the effects of KA on the cardiovascular system. These effects of KA were reduced by aminoxyacetic acid (AOAA 25 micrograms/rat, icv), an inhibitor of GABA aminotransferase. All these results described above imply that the function of GABA ergic neurons in brain can influence the effects of KA on the cardiovascular centre. We speculate that the central regulation of the cardiovascular system is involved in the interaction of GABA ergic neurons and glutamergic neurons in brain, thus, influencing the central sympathetic effects of efferent activity.


Efficacy and safety of Tanreqing injection for cough caused by acute trachea-bronchitis disease: A systematic review and meta-analysis of randomized controlled trials.
Tanreqing injection (TRQI) is an intravenous herbal preparation derived from 5 types of traditional Chinese medicines including Scutellariae Radix, Lonicerae Japonicae Flos, Forsythiae Fructus, bear bile powder and goral horn, incorporating baicalin, chlorogenic acid, ursodeoxycholic acid, and goose deoxycholic acid and other compounds known for anti-inflammatory properties, is widely used in China to treat cough caused by acute trachea-bronchitis disease (ATB). To investigate the clinical efficacy and safety of Tanreqing injection (TRQI) with and without Western medicine (WM) for cough caused by acute trachea-bronchitis (ATB). We systematically searched eight databases, including CENTRAL, Embase, PubMed, Science Direct, Wiley, China National Knowledge Infrastructure, Chinese Biomedical Literature Database and WanFang, from inception to August 2023 for randomized clinical trials (RCTs) on TRQI for cough caused by ATB. The critical outcomes of interest were time to symptom disappearance, including time for cough symptom to disappear and time to improve cough and sputum production. Important outcomes included symptom disappearance rate, adverse events (AEs) and lung function. We carried out random-effects meta-analysis using Review Manager 5.4 and assessed the certainty of evidence utilizing the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. A total of 2872 citations were identified by our search, of which 26 eligible RCTs enrolled 2731 participants. Low to moderate certainty evidence showed that when compared with WM, TRQI plus WM treatment was associated with a favorable effect on the time for cough symptom to disappear (MD -2.21 d, 95% CI -2.64 to -1.78), time to improve cough and sputum production (MD -0.68 d, 95% CI -0.83 to -0.53), symptom disappearance rate (RR 1.37, 95% CI 1.20 to 1.55), forced vital capacity, and forced expiratory volume in 1 s (MD 0.38 L, 95% CI 0.26 to 0.50; MD 2.92%, 95% CI 1.29 to 4.56, respectively). In terms of AEs, there was no association between TRQI plus WM and WM (RR 0.55, 95% CI 0.14 to 2.21; low-certainty evidence). Very low certainty evidence showed that TRQI alone was associated with reduced time to improve cough and sputum (MD -0.14 d, 95% CI -0.26 to -0.02) and increased symptom disappearance rate (RR 1.89, 95% CI 1.24 to 2.88; low certainty evidence) compared to WM. The overall efficacy of TRQI or WM for ATB cough is better than that of WM, and TRQI also effectively improve symptoms in patients with similar adverse events. However, due to the lack of methodological rigor of included studies, the present findings should be interpreted with caution. We advocate better high-quality and convincing clinical studies to be performed to prove the effectiveness and safety of TRQIs.


Bilateral macular infaction in SS disease.
A 10-year-old Afro-American female with SS disease presented with a history of painless loss of vision (RE) of 26 hours duration. Sequential fluorescein angiography revealed bilateral parafoveal occlusions and evidence of prior vaso-occlusions in each eye. Initial hydration therapy with dextrose 5% and 1/4 normal saline was followed within eight hours by transfusion with two units of packed red blood cells. Patency was restored to the occluded arterioles with return of normal acuity in LE, though the vision remains at hand motions in the RE. Sudden loss of central acuity in sickle hemoglobinopathies may result from vaso-occlusion in terminal arterioles near the foveal avascular zone. Therefore a reduced number of permanently irreversibly sickled red blood cells may prevent these macular infarcts.


Mesenchymal stem cells in arthritis: role of bone marrow microenvironment.
Based on their capacity to suppress immune responses, multipotent mesenchymal stromal cells (MSCs) are intensively studied for regenerative medicine. Moreover, MSCs are potent immunomodulatory cells that occur through the secretion of soluble mediators including nitric oxide, transforming growth factor beta, and HLAG5. The MSCs, however, are also able to express inflammatory mediators such as prostaglandin E2 or IL-6. MSCs in the bone marrow are in close contact with T cells and B cells, and they regulate immunological memory by organizing defined numbers of dedicated survival niches for plasma cells and memory T cells in the bone marrow. The role of MSCs in arthritis remains controversial - in some studies, murine allogeneic MSCs are able to decrease arthritis; in other studies, MSCs worsen the local inflammation. A recent paper in Arthritis Research and Therapy shows that bone marrow MSCs have decreased osteoblastic potential in rheumatoid arthritis, which may be related to chronic inflammation or to loss of expression of IL-1 receptor agonist. That article raises the importance of the bone marrow microenvironment for MSC biology.


Modeling the Thin-Layer Drying of Fruits and Vegetables: A Review.
The drying of fruits and vegetables is a complex operation that demands much energy and time. In practice, the drying of fruits and vegetables increases product shelf-life and reduces the bulk and weight of the product, thus simplifying transport. Occasionally, drying may lead to a great decrease in the volume of the product, leading to a decrease in storage space requirements. Studies have shown that dependence purely on experimental drying practices, without mathematical considerations of the drying kinetics, can significantly affect the efficiency of dryers, increase the cost of production, and reduce the quality of the dried product. Thus, the use of mathematical models in estimating the drying kinetics, the behavior, and the energy needed in the drying of agricultural and food products becomes indispensable. This paper presents a comprehensive review of modeling thin-layer drying of fruits and vegetables with particular focus on thin-layer theories, models, and applications since the year 2005. The thin-layer drying behavior of fruits and vegetables is also highlighted. The most frequently used of the newly developed mathematical models for thin-layer drying of fruits and vegetables in the last 10 years are shown. Subsequently, the equations and various conditions used in the estimation of the effective moisture diffusivity, shrinkage effects, and minimum energy requirement are displayed. The authors hope that this review will be of use for future research in terms of modeling, analysis, design, and the optimization of the drying process of fruits and vegetables.


Human cytomegalovirus interferes with signal transducer and activator of transcription (STAT) 2 protein stability and tyrosine phosphorylation.
We have investigated the role of signal transducer and activator of transcription (STAT) 2 during human cytomegalovirus (HCMV) replication and found that protein levels of STAT2 are downregulated. STAT2 downregulation was observed in HCMV clinical isolates and laboratory strains with the exception of strain Towne. The HCMV-induced loss of STAT2 protein occurred despite an increased accumulation of STAT2 mRNA; it required HCMV early gene expression. The decrease in STAT2 was sensitive to proteasome inhibition, suggesting degradation of STAT2 via the ubiquitin proteasome pathway. Notably, pUL27, the HCMV homologue of the mouse CMV pM27 protein, which mediates the selective proteolysis of STAT2, did not induce STAT2 downregulation. Moreover, preceding STAT2 degradation, alpha/beta interferon (IFN)-receptor-mediated tyrosine phosphorylation of STAT2 was inhibited in HCMV-infected cells. This effect was paralleled by impaired tyrosine activation of STAT1 and STAT3. Accordingly, IFNs affected the replication efficiency of STAT2 degrading and non-degrading HCMV strains to a similar degree. In summary, HCMV abrogates IFN receptor signalling at multiple checkpoints by independent mechanisms including UL27-independent degradation of STAT2 and a preceding blockade of STAT2 phosphorylation.


Targeted Delivery of Cisplatin-Derived Nanoprecursors via a Biomimetic Yeast Microcapsule for Tumor Therapy by the Oral Route.
Targeted therapy via the patient-friendly oral route remains the holy grail of chemotherapy for cancer. Herein we report a yeast-derived platform for targeted oral delivery of cisplatin (CDDP) that is one of the most effective drugs for chemotherapy of various types of cancers. Methods: The optimal conditions were first established to fabricate yeast microcapsules (YCs) with desirable loading capability. Then, CDDP-derived precursor nanoparticles (PreCDDP) were prepared and packaged into YC to produce orally deliverable PreCDDP/YC. The physiochemical properties, in vitro drug release profiles, in vitro antitumor activity, oral targeting capability, in vivo pharmacokinetics, and in vivo efficacy of the YC-based biomimetic delivery system were examined. Results: YCs obtained under the optimized condition showed desirable loading efficiency for quantum dots that were used as a model nanocargo. In vitro experiments demonstrated rapid endocytosis and prolonged retention of YC in macrophages. By electrostatic force-mediated self-deposition, PreCDDP was efficiently loaded into YC. PreCDDP/YC showed potent cytotoxicity in different tumor cells, indicating that PreCDDP loaded in YC maintained its antitumor activity after intracellular release. As compared to CDDP and PreCDDP, orally administered PreCDDP/YC displayed significantly higher bioavailability. Post oral delivery, YC could accumulate in A549 human lung carcinoma xenografts in mice, achieving by monocyte/macrophage-mediated translocation via the lymphatic system. Through this targeting effect, orally administered PreCDDP/YC showed desirable efficacy in A549 xenograft-bearing mice, which was comparable to that of free CDDP administered by intravenous injection. Orally administered free CDDP, however, did not afford antitumor effects. Furthermore, oral treatment with PreCDDP/YC displayed better safety than free CDDP administered via the oral or intravenous route. Conclusions: This biomimetic approach can serve as an effective strategy to develop targeted oral chemotherapies based on CDDP or its derivatives.


Teratoma formation of human embryonic stem cells in three-dimensional perfusion culture bioreactors.
Teratoma formation in mice is today the most stringent test for pluripotency that is available for human pluripotent cells, as chimera formation and tetraploid complementation cannot be performed with human cells. The teratoma assay could also be applied for assessing the safety of human pluripotent cell-derived cell populations intended for therapeutic applications. In our study we examined the spontaneous differentiation behaviour of human embryonic stem cells (hESCs) in a perfused 3D multi-compartment bioreactor system and compared it with differentiation of hESCs and human induced pluripotent cells (hiPSCs) cultured in vitro as embryoid bodies and in vivo in an experimental mouse model of teratoma formation. Results from biochemical, histological/immunohistological and ultrastuctural analyses revealed that hESCs cultured in bioreactors formed tissue-like structures containing derivatives of all three germ layers. Comparison with embryoid bodies and the teratomas revealed a high degree of similarity of the tissues formed in the bioreactor to these in the teratomas at the histological as well as transcriptional level, as detected by comparative whole-genome RNA expression profiling. The 3D culture system represents a novel in vitro model that permits stable long-term cultivation, spontaneous multi-lineage differentiation and tissue formation of pluripotent cells that is comparable to in vivo differentiation. Such a model is of interest, e.g. for the development of novel cell differentiation strategies. In addition, the 3D in vitro model could be used for teratoma studies and pluripotency assays in a fully defined, controlled environment, alternatively to in vivo mouse models.


Mental Health and Self-Worth in Socially Transitioned Transgender Youth.
Social transitions are increasingly common for transgender children. A social transition involves a child presenting to other people as a member of the "opposite" gender in all contexts (e.g., wearing clothes and using pronouns of that gender). Little is known about the well-being of socially transitioned transgender children. This study examined self-reported depression, anxiety, and self-worth in socially transitioned transgender children compared with 2 control groups: age- and gender-matched controls and siblings of transgender children. As part of a longitudinal study (TransYouth Project), children (9-14 years old) and their parents completed measurements of depression and anxiety (n = 63 transgender children, n = 63 controls, n = 38 siblings). Children (6-14 years old; n = 116 transgender children, n = 122 controls, n = 72 siblings) also reported on their self-worth. Mental health and self-worth were compared across groups. Transgender children reported depression and self-worth that did not differ from their matched-control or sibling peers (p = .311), and they reported marginally higher anxiety (p = .076). Compared with national averages, transgender children showed typical rates of depression (p = .290) and marginally higher rates of anxiety (p = .096). Parents similarly reported that their transgender children experienced more anxiety than children in the control groups (p = .002) and rated their transgender children as having equivalent levels of depression (p = .728). These findings are in striking contrast to previous work with gender-nonconforming children who had not socially transitioned, which found very high rates of depression and anxiety. These findings lessen concerns from previous work that parents of socially transitioned children could be systematically underreporting mental health problems.


Habituation of reinforcer effectiveness.
In this paper we propose an integrative model of habituation of reinforcer effectiveness (HRE) that links behavioral- and neural-based explanations of reinforcement. We argue that HRE is a fundamental property of reinforcing stimuli. Most reinforcement models implicitly suggest that the effectiveness of a reinforcer is stable across repeated presentations. In contrast, an HRE approach predicts decreased effectiveness due to repeated presentation. We argue that repeated presentation of reinforcing stimuli decreases their effectiveness and that these decreases are described by the behavioral characteristics of habituation (McSweeney and Murphy, 2009; Rankin etal., 2009). We describe a neural model that postulates a positive association between dopamine neurotransmission and HRE. We present evidence that stimulant drugs, which artificially increase dopamine neurotransmission, disrupt (slow) normally occurring HRE and also provide evidence that stimulant drugs have differential effects on operant responding maintained by reinforcers with rapid vs. slow HRE rates. We hypothesize that abnormal HRE due to genetic and/or environmental factors may underlie some behavioral disorders. For example, recent research indicates that slow-HRE is predictive of obesity. In contrast ADHD may reflect "accelerated-HRE." Consideration of HRE is important for the development of effective reinforcement-based treatments. Finally, we point out that most of the reinforcing stimuli that regulate daily behavior are non-consumable environmental/social reinforcers which have rapid-HRE. The almost exclusive use of consumable reinforcers with slow-HRE in pre-clinical studies with animals may have caused the importance of HRE to be overlooked. Further study of reinforcing stimuli with rapid-HRE is needed in order to understand how habituation and reinforcement interact and regulate behavior.


Assembly Dynamics and Stoichiometry of the Apoptosis Signal-regulating Kinase (ASK) Signalosome in Response to Electrophile Stress.
Apoptosis signal-regulating kinase 1 (ASK1) is a key sensor kinase in the mitogen-activated protein kinase pathway that transduces cellular responses to oxidants and electrophiles. ASK1 is regulated by a large, dynamic multiprotein signalosome complex, potentially including over 90 reported ASK1-interacting proteins. We employed both shotgun and targeted mass spectrometry assays to catalogue the ASK1 protein-protein interactions in HEK-293 cells treated with the prototypical lipid electrophile 4-hydroxy-2-nonenal (HNE). Using both epitope-tagged overexpression and endogenous expression cell systems, we verified most of the previously reported ASK1 protein-protein interactions and identified 14 proteins that exhibited dynamic shifts in association with ASK1 in response to HNE stress. We used precise stable isotope dilution assays to quantify protein stoichiometry in the ASK signalosome complex and identified ASK2 at a 1:1 stoichiometric ratio with ASK1 and 14-3-3 proteins (YWHAQ, YWHAB, YWHAH, and YWHAE) collectively at a 0.5:1 ratio with ASK1 as the main components. Several other proteins, including ASK3, PARK7, PRDX1, and USP9X were detected with stoichiometries of 0.1:1 or less. These data support an ASK signalosome comprising a multimeric core complex of ASK1, ASK2, and 14-3-3 proteins, which dynamically engages other binding partners needed to mediate diverse stress-response signaling events. This study further demonstrates the value of combining global and targeted MS approaches to interrogate multiprotein complex composition and dynamics.


Postpartum length of stay and newborn health: a cost-effectiveness analysis.
To evaluate the cost-effectiveness of increasing lengths of brief postpartum hospitalizations. A cost-effectiveness model extrapolating from secondary data was used. Social costs in 2000 US dollars were estimated using several sources, including a randomized controlled trial, a retrospective study, and survey data. Life-years saved from reduced infant mortality were estimated from administrative data from Washington State. A total of 113147 singleton newborns who were born in nonmilitary hospitals in Washington State in 1989 or 1990 and had postpartum stays short enough to be affected by length of stay legislation were studied. The cost-effectiveness of increases in postpartum lengths of stay similar to those that would occur if all mothers and singleton newborns used at least the time allotted to them under the federal length of stay legislation was measured. Estimated lower-bound cost per newborn life-year saved was 19 800 dollars (95% confidence interval: 11600-61300 dollars) when only neonatal deaths were considered. The corresponding upper-bound estimate was 94800 dollars (95% confidence interval: 55200-286800 dollars ). The results were very sensitive to assumptions about the discount rate for future life-years and the time from birth during which averted deaths are considered (neonatal deaths, postneonatal infant deaths, or all infant deaths). At hospitals that do not experience additional capacity costs as a result of increased lengths of stay, lengthening short postpartum stays seems to be more cost-effective than many common health interventions and well below cost-effectiveness thresholds suggested by the literature. Even at hospitals that experience additional capacity costs, the cost-effectiveness of lengthening short postpartum stays seems to be roughly equal to the benchmark of 100000 dollars per quality-adjusted life-year suggested by the literature.


Disease of the year: juvenile idiopathic arthritis-associated uveitis--classification and diagnostic approach.
Juvenile idiopathic arthritis (JIA) is the most common chronic rheumatic disease and the most common systemic association of uveitis in children. JIA is subdivided into seven clinically more homogeneous subtypes, using the International League of Associations for Rheumatology (ILAR) classification system: systemic arthritis, oligoarthritis, rheumatoid factor (RF)-negative polyarthritis, RF-positive polyarthritis, psoriatic arthritis, enthesitis-related arthritis, and undifferentiated arthritis. Uveitis is very rare in patients with systemic onset JIA and in patients with juvenile-onset rheumatoid arthritis. Patients with early-onset antinuclear antibody (ANA)-positive oligoarticular JIA have the highest risk of developing chronic iridocyclitis. Ocular involvement in enthesitis-related arthritis is characterized by recurrent acute symptomatic iridocyclitis. The diagnosis and differential diagnosis of JIA are based on history, physical examination, and laboratory investigations, including complete blood count, acute-phase reactants, ANA, RF, HLA-B27, and musculoskeletal imaging studies. Recent studies have revealed distinct gene expression profiles that may be useful in the diagnosis, classification, and follow-up of JIA patients.


Three-dimensional structure of ncd-decorated microtubules obtained by a back-projection method.
We have used cryo-electron microscopy and image analysis to obtain the three-dimensional (3D) structure of 11, 12, 14 and 15 protofilament microtubules decorated with the motor domain of ncd. To obtain the 3D maps, we developed a back-projection method that does not require a helical arrangement of the tubulin heterodimers. This method allows the calculation of 3D maps even when lattice discontinuities (seams) are present. The maps show that the microtubules we studied conform to a B-type lattice with one or more seams. In the presence of 5'-adenylim-idodiphosphate (AMP-PNP), the motor domain of ncd binds to the microtubule protofilament crest interacting with only one protofilament. Viewing the structures along the microtubule axis shows that the ncd motor domain and the tubulin are titled in opposite directions. We determined that a clockwise tilt of the tubulin subunits corresponds to a view from the minus end towards the plus end of the microtubule.


Tuberculosis spine: Therapeutically refractory disease.
India ranks second amongst the high-burden multi drug resistant tuberculosis (MDR-TB) countries, with an estimated incidence of 2.3% MDR-TB cases amongst the new cases and 17.2% amongst the previously treated cases. The diagnosis and treatment protocol for MDR-TB of the spine are not clearly established. We report outcome of a series of 15 cases of TB spine who were suspected to be therapeutically refractory cases (MDR-TB) on the basis of clinicoradiological failures of initial treatment. Fifteen cases of TB spine from C2 to L5 spine were suspected to be the cases of MDR-TB (therapeutically refractory cases) on the basis of failures of adequate clinicoradiological healing response at 5 months or more on antitubercular treatment (ATT). None of the patient was immunocompromised. Thirteen out of 15 patients had tissue samples sent for histopathology, culture and sensitivity, smear, BACTEC, and polymerase chain reaction (PCR). All patients were put on second line ATT and followed up fortnightly with regular liver and kidney function tests, erythrocyte sedimentation rate (ESR), and plain X-ray. Healing was documented as subjective improvement of symptoms, reduction in ESR, and observations on contrast enhanced magnetic resonance imaging (MRI) such as resolution of marrow edema, fatty replacement of bone marrow and resolution of abscesses. Ambiguous MRI observations in a few patients were resolved on positron emission tomography (PET) scan. Patients were monitored continuously for 2 years after stopping ATT. We could demonstrate a positive culture in three cases. Two of them had multi drug resistance. We could achieve healing status in 13 out of 14 patients after starting second line drugs, one patient is still on treatment while other patient with no drug resistance is responding well on ATT. The suspicion of therapeutically refractory case is of paramount importance. Once suspected, surgery to procure tissue for diagnosis and culture is to be undertaken. The demonstration of drug resistance on culture may not be achieved in all TB spine cases and empiric drug regimen for MDR-TB is to be started. We have achieved the healed status with immunomodulation and second line ATT. The length of treatment needs to be monitored with MRI and PET scan.


The antibacterial effects of ornidazole on primary molars with infected pulps.
The purpose of this study was to investigate the antibacterial effects of a short-term topical application of ornidazole on anaerobic microorganisms. The antibacterial properties of such materials against organisms at infected primary root canals have not been well documented. Twenty infected primary molars in this study were treated using ornidazole (Biteral, Roche). The bacterial contents of the roots were collected with sterile paper points before the application. Freshly mixed ornidazole and sterile saline were placed into the root canals. After 1 week, the bacterial contents of the root canals were collected again. Microbiological analyses were made. Based on our results, it appears that the antibacterial activity of ornidazole caused significant changes in rates of microorganisms (94.53% reduction).


Neonatal neurobehavioral effects of inhalation analgesia for vaginal delivery.
The authors studied the neonatal neurobehavioral effects of nitrous oxide:oxygen and enflurane:oxygen inhalation analgesia for vaginal delivery. Parturients were assigned randomly to receive no inhalation agent (Group 1, n = 21); enflurane, 0.3 to 0.8 per cent, and oxygen (Group 2, n = 22); or nitrous oxide, 30 to 50 per cent, and oxygen (Group 3, n = 18). Infants were tested at 15 min, 2 h, and 24 h of age using the Neurologic and Adaptive Capacity Score (NACS); and at 2 and 24 h using the Early Neonatal Neurobehavioral Scale (ENNS). No significant differences in neurobehavioral status occurred. For all groups, scores tended to be lowest at two hours of age. We conclude that neither enflurane nor nitrous oxide analgesia adversely affects neonatal neurobehavioral status at 15 min, 2 h, or 24 h of age.


Bone uptake of 99mTc-phosphate via labeled plasma protein.
The transfer of albumin into the extravascular space of bone and other tissues was evaluated in the non-adult rat and was found to be less rapid in bone than in the liver and kidneys but more rapid than in muscle. Protein binding of 99mTc increased continuously with time after injection of 99mTc-EHDP and -PPi. One hour after administration, about 20% of the plasma activity of 99mTc-EHDP and 90% of 99mTc-PPi was bound to proteins. Thus the possibility exists that the fixation of 99mTc through protein binding in the extravascular space of bone contributes significantly to the regular bone uptake of 99mTc-phosphates, in the event that relatively unstable compounds such as 99mTc-PPi are used.


The request to die: role for a psychodynamic perspective on physician-assisted suicide.
Published reports indicate that 2.5% of deaths in the Netherlands are the result of euthanasia or physician-assisted suicide. It is not known how many patients make these requests in the United States, but the issue has gained considerable attention, including that of the Supreme Court. The focus of the writing and discussion regarding the request to die has been on a patient's capacity. There has not been an adequate focus on the possible meanings contained within the request to die. A patient's request to die is a situation that requires the physician to engage in a dialogue to understand what the request means, including whether the request arises from a clinically significant depression or inadequately treated pain. This article outlines some of the thoughts and emotions that could underlie the patient's request to die. Recommendations are made regarding the role of the primary care physician and the role of the psychiatric consultant in the exploration of the meaning of the request.


Regulation of iron transport related genes by boron in the marine bacterium Marinobacter algicola DG893.
While there has been extensive interest in the use of boron isotope ratios as a surrogate of pH in paleoclimate studies in the context of climate change-related questions, the high (0.4 mM) concentration and the depth-independent (conservative or non-nutrient-like) concentration profile of this element have led to boron being neglected as a potentially biologically relevant element in the modern ocean. Here we report that boron affects the expression of a number of protein and genes in the "algal-associated" Gram-negative marine bacterium Marinobacter algicola DG893. Most intriguingly, a number of these proteins and genes are related to iron uptake. In a recent separate publication we have shown that boron regulates one such iron transport related protein, i.e. the periplasmic iron binding protein FbpA via a direct interaction of the metalloid with this protein. Here we show that a number of other iron uptake related genes are also affected by boron but in the opposite way i.e. they are up-regulated. We propose that the differential effect of boron on FbpA expression relative to other iron transport related genes is a result of an interaction between boron and the global iron regulatory protein Fur.


Medication adherence in HIV-infected smokers: the mediating role of depressive symptoms.
Strict medication adherence is integral to the success of highly active antiretroviral therapies (HAART) in patients with HIV. Research has examined several predictors of adherence, but few studies have examined the association between current smoking, which is highly prevalent among people living with HIV, and medication adherence; moreover, no study has examined the mediating role of depressive symptoms, which may influence both smoking and adherence. Therefore, we recruited 168 patients who were prescribed HAART and assessed viral load, CD4+ count, cigarette smoking, past week and 3-month medication adherence, and depressive symptoms. Results showed that 70% smoked at least one cigarette per day. As predicted, smoking was associated with poorer past week and 3-month adherence, and more depressive symptoms. Regression analyses provided partial support for the hypothesis that depressive symptoms mediated nonadherence among smokers. We conclude that future smoking cessation interventions with this population should target medical adherence and depression as intervention components.


Genetic Screening of Iranian Patients with 46,XY Disorders of Sex Development.
Disorders of sex development (DSDs) belong to uncommon pathologies and result from abnormalities during gonadal determination and differentiation. Various gene mutations involved in gonadal determination and differentiation have been associated with gonadal dysgenesis. Despite advances in exploration of genes and mechanisms involved in sex disorders, most children with severe 46,XY DSDs have no definitive etiological diagnoses; therefore, the possibility that other genes or loci might play important roles in these disorders needs to be explored. Patients (37) clinically suspicious for 46,XY gonadal dysgenesis (46,XY GD) of unknown etiology were studied. SRY, encoding the sex-determining region Y protein, NR5A1, encoding a transcription factor called steroidogenic factor 1, and DHH, encoding the desert hedgehog protein, were directly sequenced. Multiplex ligation-dependent probe amplification (MLPA) was used to detect deletions in NR0B1, encoding the DAX1 protein, and WNT4, encoding the WNT4 protein, and real-time PCR (qPCR) confirmed the MLPA data. Other potential loci have been investigated in the complete genome using Array-Comparative Genomic Hybridization, (Array CGH). The SRY deletion was found in five patients. One each of previously described NR5A1, DHH, and AR (androgen receptor) allelic variants were identified. A pathogenic c.2522G>A AR mutation was found in two affected brothers. A heterozygous partial deletion was found in NR5A1 and heterozygous partial duplications were found in WNT4. These deletions and duplications (del/dup) were confirmed by qPCR. The Array CGH result demonstrated one partial deletion in SOX2-OT, which encodes a member of the SOX family of transcription factors, and the exact region of the rearrangements. According to our study, del/dup mutations could be checked prior to point mutations, SOX2-OT has a potential role in gonadal dysgenesis, and Array CGH has a prominent role in gonadal dysgenesis diagnosis.


Heritability of intramammary infections at first parturition and relationships with sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield.
The objective of this study was to determine the relationships among daughter intramammary infections at first parturition and sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield. Quarter milk samples from 958 daughters (in eight Pennsylvania herds and one Nebraska herd) of 182 Holstein sires were collected within a few days of first calving and cultured to determine intramammary infection status. A total of 446 cows had intramammary infections in 835 quarters at first parturition. Incidence of intramammary infections at first parturition and the proportion of quarters infected per cow were regressed on age at first calving, days in milk at sample collection, herd-season of calving (a classification variable), and sire transmitting abilities taken one at a time. Linear effects, non-linear effects, and odds ratios were estimated for sire transmitting abilities. Separate, preplanned analyses were conducted on data from one herd that gave all heifers an intramammary antibiotic infusion in each quarter 30 d prior to the expected calving date. Separate analyses were also conducted on dependent variables that considered intramammary infections at first parturition from: all organisms, coagulase-negative staphylococci, coliform species, streptococci other than Streptococcus agalactiae, and the most common environmental organisms (coliform species and streptococci other than Streptococcus agalactiae). Daughters of sires that transmit the lowest somatic cell score had the fewest intramammary infections at first parturition. Daughters of sires that transmit longer productive life, shorter teats, and closely spaced front teats had fewer intramammary infections at first parturition. Selection for lower somatic cell score, longer productive life, shorter teats, or closely spaced front teats may reduce the incidence of intramammary infections at first parturition.


[Poststroke cognitive, emotional impairment and sleep quality: efficience of treatment with melaxen].
To study melatonin secretion and its correlations with poststroke cognitive, emotional impairment and sleep quality in the acute period of stroke and to assess treatment efficacy of melaxen. We studied 96 patients with acute stroke. A battery of tests and scales for assessment of neurological deficit, neuropsychological status and emotional impairment was used. The night urinary level of 6-sulfatoxymelatonin was assessed. The relationship between 6-sulfatoxymelatonin and cognitive, emotional status and sleep parameters was analyzed. The level of 6-sulfatoxymelatonin was decreased in the night urine. Patients with dysexecutive poststroke cognitive impairment had higher level of 6-sulfatoxymelatonin and patients with dysmnestic and mixed cognitive impairment had lower level of 6-sulfatoxymelatonin in comparison with patients with normal cognitive functions. Melaxen improved cognitive function and sleep parameters, reduced the level of anxiety in the early recovery period of stroke. A role of chronobiological processes in the development of clinical signs of stroke in the aspect of cognitive impairment is discussed.


Cytochrome a620 in Tetrahymena pyriformis. Reactions with carbon monoxide and oxygen at subzero temperatures and photochemical action spectra.
1. Mitochondria-enriched fractions of the ciliate protozoan Tetrahymena pyriformis ST contained CO-reacting cytochromes b560 and a620. 2. A non-photodissociable oxygen-containing compound of cytochrome a620 was formed in whole cell suspensions at -114 degrees C after photolysis of CO in the presence of 200 microM-O2. 3. Electron transport, indicated by the oxidation of cytochrome a620 and cytochrome c, occurred at temperatures higher than -72 degrees C. 4. Photochemical action spectra for the relief of respiratory inhibition of whole cells by CO obtained by using a liquid dye laser indicate that the only CO-reacting terminal oxidase detectable was cytochrome a620. 5. It is concluded that the alternative electron transport chains in this organism utilize non-cytochrome terminal oxidases.


Evolutionary simulations of Z-linked suppression gene drives.
Synthetic gene drives may soon be used to suppress or eliminate populations of disease vectors, pathogens, invasive species, and agricultural pests. Recent proposals have focused on using Z-linked gene drives to control species with ZW sex determination, which include Lepidopteran pests, parasitic trematodes, and cane toads. These proposals include Z-linked 'W-shredders', which would suppress populations by cleaving the W chromosome and causing females to produce only sons, as well as Z-linked female-sterilizing gene drives. Here, I use eco-evolutionary simulations to evaluate the potential of some proposed Z-linked gene drives, and to produce recommendations regarding their design and use. The simulations show that W-shredders are likely to be highly effective at eradicating populations provided that resistance to W-shredding cannot evolve. However, W-shredder alleles can invade populations from very low frequencies, making it difficult to eliminate specific populations while leaving nearby populations untouched; this issue may restrict their possible uses.


Osteoporosis and Bone Marrow Adipose Tissue.
This review focuses on the recent findings regarding bone marrow adipose tissue (BMAT) concerning bone health. We summarize the variations in BMAT in relation to age, sex, and skeletal sites, and provide an update on noninvasive imaging techniques to quantify human BMAT. Next, we discuss the role of BMAT in patients with osteoporosis and interventions that affect BMAT. There are wide individual variations with region-specific fluctuation and age- and gender-specific differences in BMAT content and composition. The Bone Marrow Adiposity Society (BMAS) recommendations aim to standardize imaging protocols to increase comparability across studies and sites. Water-fat imaging (WFI) seems an accurate and efficient alternative for spectroscopy (1H-MRS). Most studies indicate that greater BMAT is associated with lower bone mineral density (BMD) and a higher prevalence of vertebral fractures. The proton density fat fraction (PDFF) and changes in lipid composition have been associated with an increased risk of fractures independently of BMD. Therefore, PDFF and lipid composition could potentially be future imaging biomarkers for assessing fracture risk. Evidence of the inhibitory effect of osteoporosis treatments on BMAT is still limited to a few randomized controlled trials. Moreover, results from the FRAME biopsy sub-study highlight contradictory findings on the effect of the sclerostin antibody romosozumab on BMAT. Further understanding of the role(s) of BMAT will provide insight into the pathogenesis of osteoporosis and may lead to targeted preventive and therapeutic strategies.


[Azo derivatives of levorin].
Levorin, an aromatic heptaenic antibiotic and sodium nitrite in solution of acetic acid formed a diazo compound. In the diazo reaction this compound interacted with beta-napthol or H-acid. The resulting azo compounds were readily soluble in organic solvents. In water their nitrites formed true solutions. The biological activity of the azo levorins was 100-200 times lower than that of the initial levorin. The physico-chemical characteristics of the azo levorins are presented.


The cost-effectiveness of a uniform versus age-based threshold for one-off screening for prevention of cardiovascular disease.
The objective of this article was to assess the cost-effectiveness of screening strategies for cardiovascular diseases (CVD). A decision analytic model was constructed to estimate the costs and benefits of one-off screening strategies differentiated by screening age, sex and the threshold for initiating statin therapy ("uniform" or "age-adjusted") from the Spanish NHS perspective. The age-adjusted thresholds were configured so that the same number of people at high risk would be treated as under the uniform threshold. Health benefit was measured in quality-adjusted life years (QALY). Transition rates were estimated from the European Prospective Investigation into Cancer and Nutrition (EPIC-CVD), a large multicentre nested case-cohort study with 12 years of follow-up. Unit costs of primary care, hospitalizations and CVD care were taken from the Spanish health system. Univariate and probabilistic sensitivity analyses were employed. The comparator was no systematic screening program. The base case model showed that the most efficient one-off strategy is to screen both men and women at 40 years old using a uniform risk threshold for initiating statin treatment (Incremental Cost-Effectiveness Ratio of €3,274/QALY and €6,085/QALY for men and women, respectively). Re-allocating statin treatment towards younger individuals at high risk for their age and sex would not offset the benefit obtained using those same resources to treat older individuals. Results are sensitive to assumptions about CVD incidence rates. To conclude, one-off screening for CVD using a uniform risk threshold appears cost-effective compared with no systematic screening. These results should be evaluated in clinical studies.


Can enhanced recovery programmes be further improved by the addition of omega three fatty acids?
The term "enhanced recovery programme (ERP)" means applying defined protocols to augment the recovery of patients following surgery. Inflammation is body's response to insults such as infection, injury and surgical procedures. Inflammatory mediators whose function is initially protective may cause undesirable consequences, if the response is unnecessarily prolonged. The principle effects of ERP result from the reduction of the profound stress which results following major surgical procedures. A Pubmed literature search was undertaken using the keywords enhanced recovery, surgery and omega-3. The primary endpoint was whether the addition of omega-3 to ERP improved morbidity and mortality. Nine randomised trials examining the effect of omega-3 enriched diets following surgery were analysed. Inclusion of omega-3 helps in maintaining a positive nitrogen balance, overcome immune dysfunction, lower the incidence of post-operative infections with the consequence of reduced morbidity and mortality. The provision of early or continuous nutrition is one of the cornerstones of an ERP. A theoretically ideal regimen would provide an energy substrate and protein and contain a component which would limit inappropriate inflammation. The beneficial role of omega-3 results from a number of effects which limit the inflammatory response, principally by influencing the production of eicosanoids and modulating cytokines. They also enhance cell-mediated immunity and preserve immune function better than standard dietary formulations. Although ERPs have already produced significant progress, there is sufficient evidence to suggest that the provision of omega-3 fatty acids may result in further improvements.


Inflammatory rheumatologic disorders in the elderly. Unusual presentations, altered outlooks.
Comorbidities, metabolic alterations, immunosenescence, and use of drugs may affect the manifestation, clinical course, immunopathogenesis, and prognosis of inflammatory rheumatologic disease in older persons. These factors need to be considered in evaluation and treatment in the geriatric population. In this article, Drs Belostocki and Paget discuss rheumatologic disorders that typically occur in advanced age, as well as those that characteristically occur in younger patients but may present de novo in the elderly.


Vascular pericytes express osteogenic potential in vitro and in vivo.
At postconfluence, cultured bovine pericytes isolated from retinal capillaries form three-dimensional nodule-like structures that mineralize. Using a combination of Northern and Southern blotting, in situ hybridization, and immunofluorescence we have demonstrated that this process is associated with the stage-specific expression of markers of primitive clonogenic marrow stromal cells (STRO-1) and markers of cells of the osteoblast lineage (bone sialoprotein, osteocalcin, osteonectin, and osteopontin). To demonstrate that the formation of nodules and the expression of these proteins were indicative of true osteogenic potential, vascular pericytes were also inoculated into diffusion chambers and implanted into athymic mice. When recovered from the host, chambers containing pericytes were found reproducibly to contain a tissue comprised of cartilage and bone, as well as soft fibrous connective tissue and cells resembling adipocytes. This is the first study to provide direct evidence of the osteogenic potential of microvascular pericytes in vivo. Our results are also consistent with the possibility that the pericyte population in situ serves as a reservoir of primitive precursor cells capable of giving rise to cells of multiple lineages including osteoblasts, chondrocytes, adipocytes, and fibroblasts.


Role of adrenaline neurons of ventrolateral medulla (the C1 group) in the tonic and phasic control of arterial pressure.
We have sought to determine if adrenaline neurons of the C1 group are responsible for cardiovascular functions heretofore attributed to neurons in the rostral ventrolateral medulla. C1 neurons were identified in rat with antibodies to the adrenaline synthesizing enzyme, PNMT. These project to spinal cord wherein they selectively innervate the sympathetic columns. C1 neurons are also innervated by projections, mostly unilateral, from the nucleus tractus solitarii (NTS). Stimulation of the C1 area electrically, by local injection of the excitatory amino acid, L-glutamate, or with the GABA antagonist bicuculline, elevates arterial pressure (AP). Bilateral electrolytic lesions, microinjection of GABA, or administration of tetrodotoxin, in contrast, collapses AP to levels comparable to that of spinal cord transection. After lesions of one NTS, a lesion of the contralateral C1 area abolishes all reflex activity elicited by electrical or natural stimulation of baroreceptors on the side of C1 lesion without modifying resting AP. Lesions of axon bundles of PNMT neurons in the medulla also abolish baroreflexes after unilateral NTS lesions. C1 neurons appear to be the neurons mediating cardiovascular effects of application of drugs or cold to rostral portions of the ventrolateral medulla. We conclude adrenaline neurons of the C1 area represent the purportedly tonic vasomotor neurons of the rostral ventrolateral medulla and mediate the vasodepressor limb of reflexes arising from arterial baroreceptors and other cardiopulmonary afferents. Whether the tonic vasomotor response to stimulation of C1 neurons is dependent upon the release of adrenaline is not yet certain.


Novel method for monitoring genetically engineered microorganisms in the environment.
A method has been devised for directly detecting and monitoring genetically engineered microorganisms (GEMs) by using in vitro amplification of the target DNAs by a polymerase chain reaction and then hybridizing the DNAs with a specific oligonucleotide or DNA probe. A cloned 0.3-kilobase napier grass (Pennisetum purpureum) genomic DNA that did not hybridize to DNAs isolated from various microorganisms, soil sediments, and aquatic environments was inserted into a derivative of a 2,4-dichlorophenoxyacetic acid-degradative plasmid, pRC10, and transferred into Escherichia coli. This genetically altered microorganism, seeded into filter-sterilized lake and sewage water samples (10(4)/ml), was detected by a plate count method in decreasing numbers for 6 and 10 days of sample incubation, respectively. The new method detected the amplified unique marker (0.3-kilobase DNA) of the GEM even after 10 to 14 days of incubation. This method is highly sensitive (it requires only picogram amounts of DNA) and has an advantage over the plate count technique, which can detect only culturable microorganisms. The method may be useful for monitoring GEMs in complex environments, where discrimination between GEMs and indigenous microorganisms is either difficult or requires time-consuming tests.


Induced phase transformation in ionizable colloidal nanoparticles.
Acid-base equilibria directly influence the functionality and behavior of particles in a system. Due to the ionizing effects of acid-base functional groups, particles will undergo charge exchange. The degree of ionization and their intermolecular and electrostatic interactions are controlled by varying the pH and salt concentration of the solution in a system. Although the pH can be tuned in experiments, it is hard to model this effect using simulations or theoretical approaches. This is due to the difficulty in treating charge regulation and capturing the cooperative effects in a colloidal suspension with Coulombic interaction. In this work, we analyze a suspension of ionizable colloidal particles via molecular dynamics (MD) simulations, along with Monte Carlo simulations for charge regulation (MC-CR) and derive a phase diagram of the system as a function of pH. It is observed that as pH increases, particles functionalized with acid groups change their arrangement from face-centered cubic (FCC) packing to a disordered state. We attribute these transitions to an increase in the degree of charge polydispersity arising from an increase in pH. Our work shows that charge regulation leads to amorphous solids in colloids when the mean nanoparticle charge is sufficiently high.


Protein palmitoylation by a family of DHHC protein S-acyltransferases.
Protein palmitoylation refers to the posttranslational addition of a 16 carbon fatty acid to the side chain of cysteine, forming a thioester linkage. This acyl modification is readily reversible, providing a potential regulatory mechanism to mediate protein-membrane interactions and subcellular trafficking of proteins. The mechanism that underlies the transfer of palmitate or other long-chain fatty acids to protein was uncovered through genetic screens in yeast. Two related S-palmitoyltransferases were discovered. Erf2 palmitoylates yeast Ras proteins, whereas Akr1 modifies the yeast casein kinase, Yck2. Erf2 and Akr1 share a common sequence referred to as a DHHC (aspartate-histidine-histidine-cysteine) domain. Numerous genes encoding DHHC domain proteins are found in all eukaryotic genome databases. Mounting evidence is consistent with this signature motif playing a direct role in protein acyltransferase (PAT) reactions, although many questions remain. This review presents the genetic and biochemical evidence for the PAT activity of DHHC proteins and discusses the mechanism of protein-mediated palmitoylation.


Jumping the barrier to beta-lactam resistance in Staphylococcus aureus.
Although the staphylococcal methicillin resistance determinant, mecA, resides on a mobile genetic element, staphylococcus cassette chromosome mec (SCCmec), its distribution in nature is limited to as few as five clusters of related methicillin-resistant Staphylococcus aureus (MRSA) clones. To investigate the potential role of the host chromosome in clonal restriction of the methicillin resistance determinant, we constructed plasmid pYK20, carrying intact mecA, and introduced it into several methicillin-susceptible Staphylococcus aureus strains, five of which were naive hosts (i.e., mecA not previously resident on the host chromosome) and five of which were experienced hosts (i.e., methicillin-susceptible variants of MRSA strains from which SCCmec was excised). We next assessed the effect of the recipient background on the methicillin resistance phenotype by population analysis, by assaying the mecA expression of PBP2a by Western blot analysis, and by screening for mutations affecting mecA. Each experienced host transformed with pYK20 had a resistance phenotype and expressed PBP2a similar to that of the parent with chromosomal SCCmec, but naive hosts transformed with pYK20 selected against its expression, indicative of a host barrier. Either inducible beta-lactamase regulatory genes blaR1-blaI or homologous regulatory genes mecR1-mecI, which control mecA expression, acted as compensatory elements, permitting the maintenance and expression of plasmid-carried mecA.


Frequency of sit-down patient care rounds, attainment of clinical performance targets, hospitalization, and mortality in hemodialysis patients.
Sit-down patient rounding in hemodialysis units allows providers to focus collectively on each patient's needs and may affect patient outcomes positively. The objective was to examine whether sit-down rounding practices improve patient outcomes in a cohort of 644 adult hemodialysis patients from 75 outpatient dialysis clinics in 17 states throughout the United States who survived at least 6 mo after enrollment (average follow-up, 3.2 yr). Achievement of well-accepted 6-mo clinical performance targets of albumin (> or =3.5 g/dl), hemoglobin (> or =11 g/dl), calcium-phosphate product (<60 mg(2)/dl(2)), dose (Kt/V > or =1.2), and vascular access type (fistula); hospitalization rates; and all-cause mortality served as outcomes. Monthly or more frequent sit-down rounds were conducted in 36 (48%) of 75 clinics, representing 287 (45%) of 644 patients. More frequent sit-down rounds were positively associated with an increased chance of achieving the 6-mo clinical performance target for albumin compared with less frequent rounds (odds ratio [OR], 1.88; 95% confidence interval [CI], 1.12 to 3.15); patients who were treated at clinics with more frequent rounds also had nearly twice the odds of achieving more of the five performance targets (OR, 1.95; 95% CI, 1.11 to 3.42). After adjustment for potential confounders, patients who were treated at clinics with more frequent sit-down rounds were 32% less likely to be hospitalized (incidence rate ratio, 0.68; 95% CI, 0.51 to 0.91), had fewer hospital days per year (rate ratio, 0.50; 95% CI, 0.26 to 0.98), and were 29% less likely to die (relative hazard, 0.71; 95% CI, 0.53 to 0.95). Adjustment for some clinical performance targets attenuated the statistical significance of the association with hospitalization. More frequent sit-down rounds in hemodialysis units are associated with better patient outcomes, including an increased chance of meeting the albumin clinical performance target, decreased hospitalization, and decreased risk of mortality. This association may be due to the positive effect of collaborative discussion by the patient care team of short- and long-term care goals for individual patients.


Role of Vitamin D in Inflammatory Bowel Disease.
Vitamin D is a secosteroid hormone that possesses immunomodulatory properties and has been demonstrated to potentially influence inflammatory bowel disease (IBD) pathogenesis and activity. Epidemiologic data have associated vitamin D deficiency with an increased risk of IBD, hospitalizations, surgery, and loss of response to biologic therapy. Conversely, IBD itself can lead to vitamin D deficiency. This bidirectional relationship between vitamin D and IBD suggests the need for monitoring and repletion of vitamin D, as needed, in the IBD patient. This review discusses the role of vitamin D in IBD and provides practical guidance on vitamin D repletion.


Mitochondrial DNA transmitted from sperm in the blue mussel Mytilus galloprovincialis showing doubly uniparental inheritance of mitochondria, quantified by real-time PCR.
Doubly uniparental inheritance (DUI) of mitochondrial DNA transmission to progeny has been reported in the mussel, Mytilus. In DUI, males have both paternally (M type) and maternally (F type) transmitted mitochondrial DNA (mtDNA), but females have only the F type. To estimate how much M type mtDNA enters the egg with sperm in the DUI system, ratios of M type to F type mtDNA were measured before and after fertilization. M type mtDNA content in eggs increased markedly after fertilization. Similar patterns in M type content changes after fertilization were observed in crosses using the same males. To compare mtDNA quantities, we subsequently measured the ratios of mtDNA to the 28S ribosomal RNA gene (an endogenous control sequence) in sperm or unfertilized eggs using a real-time polymerase chain reaction (PCR) assay. F type content in unfertilized eggs was greater than the M type in sperm by about 1000-fold on average. M type content in spermatozoa was greater than in unfertilized egg, but their distribution overlapped. These results may explain the post-fertilization changes in zygotic M type content. We previously demonstrated that paternal and maternal M type mtDNAs are transmitted to offspring, and hypothesized that the paternal M type contributed to M type transmission to the next generation more than the maternal type did. These quantitative data on M and F type mtDNA in sperm and eggs provide further support for that hypothesis.


Detection of intergenerational genetic effects with application to HLA-B matching as a risk factor for schizophrenia.
Association studies using unrelated individuals cannot detect intergenerational genetic effects contributing to disease. To detect these effects, we improve the extended maternal-fetal genotype (EMFG) incompatibility test to estimate any combination of maternal effects, offspring effects, and their interactions at polymorphic loci or multiple SNPs, using any size pedigrees. We explore the advantages of using extended pedigrees rather than nuclear families. We apply our methods to schizophrenia pedigrees to investigate whether the previously associated mother-daughter HLA-B matching is a genuine risk or the result of bias. Simulations demonstrate that using the EMFG test with extended pedigrees increases power and precision, while partitioning extended pedigrees into nuclear families can underestimate intergenerational effects. Application to actual data demonstrates that mother-daughter HLA-B matching remains a schizophrenia risk factor. Furthermore, ascertainment and mate selection biases cannot by themselves explain the observed HLA-B matching and schizophrenia association. Our results demonstrate the power of the EMFG test to examine intergenerational genetic effects, highlight the importance of pedigree rather than case/control or case-mother/control-mother designs, illustrate that pedigrees provide a means to examine alternative, non-causal mechanisms, and they strongly support the hypothesis that HLA-B matching is causally involved in the etiology of schizophrenia in females.


A multidimensional and hierarchical carbon-confined cobalt phosphide nanocomposite as an advanced anode for lithium and sodium storage.
Transition metal phosphides possess remarkable theoretical charge capacities; however, they demonstrate poor intrinsic electrical conductivity and enormous structure changes during cycling. Herein, a novel, multidimensional and hierarchical nanostructure is synthesized which constitutes 0D CoP nanoparticles distrbuted in a 1D dual carbon matrix and further homogenously encapsulated in a 3D graphene (GR) network; this material is denoted as CoP@DC@GR. This synergistic design produces superior advantages for lithium and sodium storage: (i) the ultrasmall and highly dispersed 0D CoP nanoparticles embedded in the 1D dual carbon matrix can decrease the electron/ion transport paths; (ii) the 3D interconnected architecture constructed from 2D GR and the 1D dual carbon matrix guarantees a robust structure to withstand the mechanical stress of the 1D dual carbon matrix, which can relieve volume inflation of the 0D CoP nanoparticles; (iii) the 1D dual carbon matrix and 3D GR frame structure offer continuous electron/ion transport routes, thus promoting rapid reaction kinetics. Due to this multiscale coordinated design, the CoP@DC@GR nanocomposite demonstrates high charge capacity, remarkable cycle stability, and distinguished rate performance; therefore, it possesses fascinating potential in anodes for lithium and sodium storage.


A C 21 -Steroidal Glycoside Isolated from the Roots of Cynanchum auriculatum Induces Cell Cycle Arrest and Apoptosis in Human Gastric Cancer SGC-7901 Cells.
Caudatin 3-O-β-D-cymaropyranosyl-(1 → 4)-β-D-oleandropyranosyl-(1 → 4)-β-D-cymaropyranosyl-(1 → 4)-β-D-cymaropyranoside (CGII) is one of the C21-steroidal glycosides isolated from the roots of Cynanchum auriculatum ROYLE ex WIGHT. This study aimed to determine the cell growth, cell proliferation, and apoptotic cell death of human gastric cancer cells after CGII treatment. MTT assay was used to determine cell growth; fluorescence-activated cell sorting analysis was used to evaluate cell cycle distribution and apoptotic cell death. Immunoblotting was applied for measuring the expression of proteins involved in the cell cycle progression. The activities of caspase-3, -8, and -9 were detected by colorimetric caspase activity assays. CGII inhibited cell growth of human gastric cancer SGC-7901 cells in a concentration- and time-dependent manner. Treatment of SGC-7901 cells with CGII resulted in G1 phase cell cycle arrest, accompanied with decreased expression of cyclin D1 and cyclin-dependent kinases 4 and 6. CGII induced cell apoptosis and activated caspase-3, caspase-8, and caspase-9. In contrast, pan-caspase inhibitor z-VAD-fmk partially abolished the CGII-induced growth inhibition of SGC-7901 cells. In conclusion, CGII inhibits cell growth of human gastric cancer cells by inducing G1 phase cell cycle arrest and caspase-dependent apoptosis cascades.


Particle attached and free floating pathogens survival kinetics under typical stream and thermal spring temperature conditions.
Improved understanding of pathogen survival in the stream environment is needed to enhance existing predictive models of stream pathogen populations. Further, the increasing use of thermal springs for bathing necessitates additional studies focused on not only typical streams but also thermal spring conditions, where water temperature is relatively higher than typical streams. This study was conducted to assess the survival of E. coli O157:H7 and Salmonella Typhimurium in stream water under free floating and particle-attached conditions at a range of temperature. A series of microcosm studies were conducted to determine pathogen decay rates. In bench-scale experiments, water circulation and sediment resuspension mimicked natural stream and thermal spring conditions, with continuous air flow providing aeration, constant mixing and turbulent conditions, and improved water circulation. Data on E. coli O157:H7 and Salmonella survival were subsequently used to determine first-order decay equations for calculating the rate constant and decimal reduction time for the modeled experimental conditions. Results showed that at 40 °C, the survival of particle attached E. coli O157:H7 was longer than that of particle attached Salmonella. Under free floating condition, Salmonella survived longer than E. coli O157:H7. At 50 °C, survival of particle attached E. coli O157:H7 and Salmonella was longer than that of free floating E. coli and Salmonella. At 60 °C, survival of particle attached Salmonella was longer than that of free floating Salmonella. Similarly at 60 °C, the survival of E. coli O157:H7 under particle attached condition was longer than that of the free floating condition. The findings of this study suggest that the survival of E. coli O157:H7 differs than the survival of Salmonella in stream water and thermal spring conditions, and the assumption used in previous studies to estimate survival of bacteria in stream environment could result in over/underestimation if the impact of particle attachment on pathogen survival is not accounted for.


6-year change in high sensitivity cardiac troponin T and the risk of atrial fibrillation in the Atherosclerosis Risk in Communities cohort.
Circulating high sensitivity cardiac troponin T (hs-cTnT) is associated with incidence of atrial fibrillation (AF), but the association of changes in hs-cTnT over time on incident AF has not been explored. Six-year increase in circulating hs-cTnT will be associated with increased risk of AF and will contribute to improved prediction of incident AF. We conducted a prospective cohort analysis of 8431 participants from the Atherosclerosis Risk in Communities (ARIC) study. hs-cTnT change was categorized at visit 2 and 4 as undetectable (<5 ng/L), detectable (≥5 ng/L, <14 ng/L), or elevated (≥14 ng/L). We used Cox regression to examine the association between the combination of hs-cTnT categories at two visits and incident AF. We also assessed the impact of adding absolute hs-cTnT change on risk discrimination for AF by C-statistics and net reclassification improvement (NRI). Over a mean follow-up of 16.5 years, 1629 incident AF cases were diagnosed. Among participants with undetectable hs-cTnT at visit 2, the multivariable HR of AF was 1.28 (95% CI 1.12-1.48) among those with detectable or elevated hs-cTnT at visit 4 compared to those in which hs-cTnT remained undetectable. Among those with detectable hs-cTnT at visit 2, compared to those who remained in the detectable hs-cTnT group, reduction to undetectable at visit 4 was associated with lower risk of AF (HR 0.74, 95% CI 0.59-0.94), while increment to elevated was associated with higher AF risk (HR 1.30, 95% CI 1.01-1.68). Adding hs-cTnT change to our main model with baseline hs-cTnT did not result in significant improvement in the C-statistic or substantial NRI. Six-year increase in circulating hs-cTnT was associated with elevated risk of incident AF.


Lifestyle counseling in patients with carotid arteriosclerosis from Luxemburg should focus more on the reduction of sugar, sodium and saturated fat consumption.
Healthy diet and physical activity improve risk factors for cerebrovascular disease. It is unclear whether patients with carotid artery disease from Luxemburg meet common guideline criteria and whether systematic counseling has a sustained effect. We assessed anthropometric data, eating habits and physical activity habits in 53 patients with carotid atherosclerosis at baseline, after 4 and 20 weeks, and advised them five times for 30 min to follow a modified Mediterranean diet and to perform moderate physical exercise at least during 30 min/day. The patients had a mildly increased BMI (mean 27.6, recommended below 25), they already ate enough vegetables and fruits (mean 485 g daily, recommended at least 400 g), they ate too much sugar (mean 74 g daily) and sodium (mean 2710 mg daily, recommended less than 1500), they consumed 13% of calories from saturated fatty acids (recommended less than 10%), and they already moved sufficiently (62 min daily of moderate and intense physical activity, recommended at least 30 min of moderate physical activity). Lifestyle counseling had a sustained effect on weight, reduction of global caloric intake, carbohydrate and cholesterol intake and on an increase in consumption of poly-unsaturated fatty acids, vegetables and fibres. There was no sustained effect on the consumption of sugar, sodium, and saturated fat. The reduction of sugar, sodium and saturated fat consumption should be stressed more in counselling of this patient group.


Epigenetic regulation of embryonic stem cell marker miR302C in human chondrosarcoma as determinant of antiproliferative activity of proline-rich polypeptide 1.
Metastatic chondrosarcoma of mesenchymal origin is the second most common bone malignancy and does not respond either to chemotherapy or radiation; therefore, the search for new therapies is relevant and urgent. We described recently that tumor growth inhibiting cytostatic proline-rich polypeptide 1, (PRP-1) significantly upregulated tumor suppressor miRNAs, downregulated onco-miRNAs in human chondrosarcoma JJ012 cell line, compared to chondrocytes culture. In this study we hypothesized the existence and regulation of a functional marker in cancer stem cells, correlated to peptides antiproliferative activity. Experimental results indicated that among significantly downregulated miRNA after PRP-1treatment was miRNAs 302c*. This miRNA is a part of the cluster miR302‑367, which is stemness regulator in human embryonic stem cells and in certain tumors, but is not expressed in adult hMSCs and normal tissues. PRP-1 had strong inhibitory effect on viability of chondrosarcoma and multilineage induced multipotent adult cells (embryonic primitive cell type). Unlike chondrosarcoma, in glioblastoma, PRP-1 does not have any inhibitory activity on cell proliferation, because in glioblastoma miR-302-367 cluster plays an opposite role, its expression is sufficient to suppress the stemness inducing properties. The observed correlation between the antiproliferative activity of PRP-1 and its action on downregulation of miR302c explains the peptides opposite effects on the upregulation of proliferation of adult mesenchymal stem cells, and the inhibition of the proliferation of human bone giant-cell tumor stromal cells, reported earlier. PRP-1 substantially downregulated the miR302c targets, the stemness markers Nanog, c-Myc and polycomb protein Bmi-1. miR302c expression is induced by JMJD2-mediated H3K9me2 demethylase activity in its promoter region. JMJD2 was reported to be a positive regulator for Nanog. Our experimental results proved that PRP-1 strongly inhibited H3K9 activity comprised of a pool of JMJD1 and JMJD2. We conclude that inhibition of H3K9 activity by PRP-1 leads to downregulation of miR302c and its targets, defining the PRP-1 antiproliferative role.


Structural features and biological activities of the cathelicidin-derived antimicrobial peptides.
Cathelicidins are a numerous group of mammalian proteins that carry diverse antimicrobial peptides at the C-terminus of a highly conserved preproregion. These peptides, which become active when released from the proregion, display a remarkable variety of sizes, sequences, and structures, and in fact comprise representatives of all the structural groups in which the known antimicrobial peptides have been classified. Most of the cathelicidin-derived peptides exert a broad spectrum and potent antimicrobial activity and also bind to lipopolysaccharide and neutralize its effects. In addition, some of them have recently been shown to exert other activities and might participate in host defense also by virtue of their ability to induce expression of molecules involved in a variety of biological processes. This review is aimed at providing a general overview of the cathelicidins and of the peptides derived therefrom, with emphasis on aspects such as structure, biological activities in vitro and in vivo, and structure/activity relationship studies.


Frontal linear scleroderma (en coup de sabre).
En coup de sabre is a type of linear scleroderma that presents on the frontal or frontoparietal scalp. The case of a 54-year-old woman with a 3 and one-half year history of en coup de sabre is presented. The clinical presentation, laboratory findings, epidemiology, etiologic hypotheses, and treatment options are reviewed.


Implications of diadochokinesia in children with speech sound disorder.
To verify the performance of children with and without speech sound disorder in oral motor skills measured by oral diadochokinesia according to age and gender and to compare the results by two different methods of analysis. Participants were 72 subjects aged from 5 years to 7 years and 11 months divided into four subgroups according to the presence of speech sound disorder (Study Group and Control Group) and age (<6 years and 5 months and >6 years and 5 months). Diadochokinesia skills were assessed by the repetition of the sequences 'pa', 'ta', 'ka' and 'pataka' measured both manually and by the software Motor Speech Profile®. Gender was statistically different for both groups but it did not influence on the number of sequences per second produced. Correlation between the number of sequences per second and age was observed for all sequences (except for 'ka') only for the control group children. Comparison between groups did not indicate differences between the number of sequences per second and age. Results presented strong agreement between the values of oral diadochokinesia measured manually and by MSP. This research demonstrated the importance of using different methods of analysis on the functional evaluation of oro-motor processing aspects of children with speech sound disorder and evidenced the oro-motor difficulties on children aged under than eight years old.


Cutting edge: TNFR-shedding by CD4+CD25+ regulatory T cells inhibits the induction of inflammatory mediators.
CD4+CD25+ regulatory T (Treg) cells play an essential role in maintaining tolerance to self and nonself. In several models of T cell-mediated (auto) immunity, Treg cells exert protective effects by the inhibition of pathogenic T cell responses. In addition, Treg cells can modulate T cell-independent inflammation. We now show that CD4+CD25+ Treg cells are able to shed large amounts of TNFRII. This is paralleled by their ability to inhibit the action of TNF-alpha both in vitro and in vivo. In vivo, Treg cells suppressed IL-6 production in response to LPS injection in mice. In contrast, Treg cells from TNFRII-deficient mice were unable to do so despite their unhampered capacity to suppress T cell proliferation in a conventional in vitro suppression assay. Thus, shedding of TNFRII represents a novel mechanism by which Treg cells can inhibit the action of TNF, a pivotal cytokine driving inflammation.


Herpes simplex virus type 1 latency-associated transcript inhibits apoptosis and promotes neurite sprouting in neuroblastoma cells following serum starvation by maintaining protein kinase B (AKT) levels.
The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) is expressed abundantly in latently infected sensory neurons. LAT-deletion-mutant virus strains have reduced-reactivation phenotypes in small animal models of infection, demonstrating that LAT plays an important role in the latency-reactivation cycle of HSV-1. Previous studies demonstrated that the anti-apoptosis functions of LAT are important for regulating the latency-reactivation cycle because three different anti-apoptosis genes can substitute for LAT. Although LAT inhibits caspase 3 activation, the signalling pathway by which LAT inhibits caspase 3 activation was not identified. In this study, we analysed mouse neuroblastoma cells (C1300) that express LAT stably (DC-LAT6 cells) following serum starvation. As expected, DC-LAT6 cells were resistant to apoptosis following serum withdrawal. Levels of total and phosphorylated AKT (protein kinase B), a serine/threonine protein kinase that promotes cell survival, were higher in DC-LAT6 cells after serum withdrawal than in C1300 cells or a cell line stably transfected with a LAT promoter mutant (DC-DeltaLAT311). A specific AKT inhibitor reduced the anti-apoptosis functions of LAT and phosphorylated AKT levels. After serum withdrawal, more DC-LAT6 cells sprouted neurites and exhibited a differentiated morphology. NeuN (neuronal nuclei), a neuron-specific nuclear protein, was expressed abundantly in DC-LAT6 cells, but not C1300 cells, after serum withdrawal, further supporting the concept that LAT enhanced neuronal-like morphology. Collectively, these studies suggested that LAT, directly or indirectly, maintained total and phosphorylated AKT levels, which correlated with increased cell survival and mature neuronal-like morphology.


Equity and geography: the case of child mortality in Papua New Guinea.
Recent assessments show continued decline in child mortality in Papua New Guinea (PNG), yet complete subnational analyses remain rare. This study aims to estimate under-five mortality in PNG at national and subnational levels to examine the importance of geographical inequities in health outcomes and track progress towards Millennium Development Goal (MDG) 4. We performed retrospective data validation of the Demographic and Health Survey (DHS) 2006 using 2000 Census data, then applied advanced indirect methods to estimate under-five mortality rates between 1976 and 2000. The DHS 2006 was found to be unreliable. Hence we used the 2000 Census to estimate under-five mortality rates at national and subnational levels. During the period under study, PNG experienced a slow reduction in national under-five mortality from approximately 103 to 78 deaths per 1,000 live births. Subnational analyses revealed significant disparities between rural and urban populations as well as inter- and intra-regional variations. Some of the provinces that performed the best (worst) in terms of under-five mortality included the districts that performed worst (best), with district-level under-five mortality rates correlating strongly with poverty levels and access to services. The evidence from PNG demonstrates substantial within-province heterogeneity, suggesting that under-five mortality needs to be addressed at subnational levels. This is especially relevant in countries, like PNG, where responsibility for health services is devolved to provinces and districts. This study presents the first comprehensive estimates of under-five mortality at the district level for PNG. The results demonstrate that for countries that rely on few data sources even greater importance must be given to the quality of future population surveys and to the exploration of alternative options of birth and death surveillance.


Recurrent moderate hypoglycemia exacerbates oxidative damage and neuronal death leading to cognitive dysfunction after the hypoglycemic coma.
Moderate recurrent hypoglycemia (RH) is frequent in Type 1 diabetes mellitus (TIDM) patients who are under intensive insulin therapy increasing the risk for severe hypoglycemia (SH). The consequences of RH are not well understood and its repercussions on neuronal damage and cognitive function after a subsequent episode of SH have been poorly investigated. In the current study, we have addressed this question and observed that previous RH during seven consecutive days exacerbated oxidative damage and neuronal death induced by a subsequent episode of SH accompanied by a short period of coma, in the parietal cortex, the striatum and mainly in the hippocampus. These changes correlated with a severe decrease in reduced glutathione content (GSH), and a significant spatial and contextual memory deficit. Administration of the antioxidant, N-acetyl-L-cysteine, (NAC) reduced neuronal death and prevented cognitive impairment. These results demonstrate that previous RH enhances brain vulnerability to acute hypoglycemia and suggests that this effect is mediated by the decline in the antioxidant defense and oxidative damage. The present results highlight the importance of an adequate control of moderate hypoglycemic episodes in TIDM.


Mycobacterium indicus pranii therapy induces tumor regression in MyD88- and TLR2-dependent manner.
Mycobacterium indicus pranii (MIP) is an atypical mycobacterium species with potent antitumor efficacy. Macrophages and dendritic cells (DCs) are antigen-presenting cells, playing key roles in the activation of antitumor immunity. We have previously shown the potent activation of macrophages and DCs by MIP, which is mediated by MyD88-TLR2 signaling axis. In the present study, we further examined the role of MyD88 and TLR2 in MIP-mediated tumor regression. Wild-type and MyD88-/- mice were implanted with B16F10 tumor cells, treated with MIP or phosphate-buffered saline (PBS) and monitored for tumor growth. As expected, MIP therapy led to significant tumor regression in wild-type mice. However, antitumor efficacy of MIP was lost in MyD88-/- animals. Both PBS-treated (control) and MIP-treated MyD88-/- mice developed tumors with comparable volume. Since MyD88 relays TLR engagement signals, we analyzed the antitumor efficacy of MIP in TLR2-/- and TLR4-/- mice. It was observed that MIP therapy reduced tumor burden in wild-type and TLR4-/- mice but not in TLR2-/- mice. Tumor volume in MIP-treated TLR2-/- mice were comparable with those in PBS-treated wild-type animals. These results implicated the MyD88-TLR2 signaling axis in the antitumor efficacy of MIP.