[Effect of emodin on NO-cGMP signal pathway in rat vascular endothelium in vitro].
To investigate the vasorelaxation effect of emodin and its relationship with NO-cGMP signal pathway. Changes of tension of rat thoracic aortic rings were measured by MedLab biologic signal collection system, and the activity of total nitric oxide synthase (tNOS), constitutive NOS (cNOS) and inducible NOS (iNOS) in endothelium after being treated with emodin was determined with nitric acid reductase method. Emodin relaxed the phenylephrine and potasium chlorate induced contraction of aortic rings, either with or without intact endothelium, in a concentration-dependent manner. Pretreatment of no-specific potassium channel blocker strontium chloride (CsCL) could attenuate the vasorelaxation effect of emodin on aortic rings without intact endothelium, but it could not inhibit vasorelaxation of emodin on aortic rings with intact endothelium. This vasorelaxation action of emodin (40 micromol/L) could be partial blocked by NOS inhibitor L-NAME and guanylate cyclase inhibitor ODQ, with the vasorelaxation range dropped to 64.76 +/- 13.73% and 6.28 +/- 4.79% respectively. Moreover, emodin (40 micromol/L) increased iNOS activity significantly. The concentration-dependent vasorelaxation effect of emodin might act by activating the NO-cGMP pathway in vascular endothelium.


Idiopathic or primary windswept deformity: the etiological significance of the radiological findings.
The radiographs of 16 knees in eight children who had a valgus deformity of one knee in association with a varus deformity in the other knee were studied. The condition, idiopathic or primary windswept deformity, occurs in 1- to 3-year-old healthy children with normal developmental milestones. The onset of radiological and clinical deformity is abrupt. The disease is a physeal osteochondrosis, and two varieties of it are identical to Blount's tibia vara and to tibia valga, respectively.


IgG4-related kidney disease complicated with retroperitoneal fibrosis: A case report.
IgG4-related disease (IgG4-RD) is a chronic fibrotic disease mediated by immunity recognized by clinicians in recent years. When the kidney is involved, it is called IgG4-related kidney disease (IgG4-RKD). IgG4-related tubulointerstitial nephritis (IgG4-TIN) is a representative manifestation of IgG4-RKD. IgG4-TIN can cause obstructive nephropathy complicated by retroperitoneal fibrosis (RPF). Cases of IgG4-TIN complicated with RPF are rare. Glucocorticoids are the first-line therapeutic medication for IgG4-RD and can significantly improve renal function. Herein, we report the case of a 56-year-old man with IgG4-RKD complicated with RPF. The patient presented to the hospital with complaints of elevated serum creatinine (Cr), nausea, and vomiting. During hospitalization, Cr was 1448.6 µmol/L, and serum IgG4 was increased. A total abdominal computed tomography (CT) scan and enhanced CT scan obviously indicated RPF. Although this patient had a long course and renal insufficiency, we performed a kidney biopsy. Renal biopsy showed that the renal tubulointerstitium had focal plasma cell infiltration and increased lymphocyte infiltration accompanied by fibrosis. After combining the biopsy results with immunohistochemistry, it was found that the absolute number of positive IgG4+ cells per high power field exceeded 10, and the ratio of IgG4/IgG was over 40%. Finally, the patient was diagnosed with IgG4-TIN complicated with RPF and given glucocorticoids as long-term maintenance therapy, helping him keep out of dialysis. After a follow-up of 19 mo, the patient had recovered well. Previous literature on IgG4-RKD and RPF was retrieved from PubMed to characterize the clinical and pathological features and to identify the diagnosis and treatment of IgG4-RKD. Our case report demonstrates the clinical characteristics of IgG4-RKD complicated with RPF. Serum IgG4 is a favorable indicator for screening. Performing renal biopsy actively plays a vital role in diagnosis and treatment, even if the patient has a long course and manifests with renal insufficiency. It is remarkable to treat IgG4-RKD with glucocorticoids. Hence, early diagnosis and targeted therapy are essential for reversing renal function and improving extrarenal manifestations in patients with IgG4-RKD.


Reproductive history in women with coronary heart disease. A case-control study.
Reproductive history events may be risk factors for coronary heart disease among women under age 60. In this study, the cases were 169 women under age 60 with a diagnosis of coronary heart disease while residents of Rochester, Minnesota. The first manifestation of coronary heart disease was angina in 95, myocardial infarction in 59, and sudden unexpected death in 15 women. Two age- and sex-matched controls for each case were selected from all registrations for care at the Mayo Clinic in the same year the case was diagnosed. The cases had more of the accepted coronary heart disease risk factors--i.e., hypertension (relative risk = 2.9), smoking (relative risk = 2.4), elevated serum cholesterol (relative risk = 2.6), and diabetes (relative risk = 4.1)--than did controls. In addition, women undergoing artificial menopause and those whose first pregnancy was before age 25 appeared to be at higher risk of coronary heart disease. Use of multivariate analysis to control for known coronary heart disease risk factors resulted in a relative risk of 1.9 for women whose age at first pregnancy was less than 20, and 1.8 for those whose age at first pregnancy was 20-24, as compared with those never pregnant and those whose age at first pregnancy was greater than or equal to 25.


Comparative bone marrow clastogenicity of cigarette sidestream, mainstream and recombined smoke condensates in mice.
The chromosome-damaging effects of cigarette sidestream (SS) and mainstream (MS) smoke condensates and a mixture of these were compared in 8-week-old NMRI mice by intraperitoneal administration. Each filtered commercial brand of cigarette was smoked by a smoking machine under the standard conditions, and the separately collected SS and MS smoke condensates were extracted with acetone/methanol as described elsewhere. The extracts were tested before and after treatment of animals with an enzyme inducer (Aroclor 1254) or inhibitor (Metyrapone). Increased formation of micronuclei within polychromatic erythrocytes (PCEs) of femural bone marrow 30 h after injection of the extracts was regarded as being due to a clastogenic effect. Regardless of the type of smoke extract injected, the increased formation of micronuclei was found to be dose dependent. The SS smoke condensate induced approximately 29% more micronuclei than the MS smoke condensate, the difference being significant (P less than 0.01). The overall clastogenicity of a 1:1 mixture of SS and MS smoke condensates was not substantially different from the activity of either SS or MS smoke condensate alone. Pretreatment of animals with Aroclor clearly enhanced the differences between the number of micronucleated PCEs caused by SS versus MS smoke condensate; SS smoke condensate induced 50% more micronuclei than did MS smoke condensate (P less than 0.001). Pretreatment of mice with Metyrapone did not modify appreciably the induction of micronuclei by either type of smoke. These results are discussed with reference to our previous data involving inhalation experiments and the recent issue of passive smoking.


Sequential observation of gastric ulcer healing by endoscopic ultrasonography.
The relationship between superficial findings and the inner structure of gastric ulcers is unclear. The present study was undertaken to clarify the relationship between them. In the first study, 43 patients with active gastric ulcers were examined by endoscopic ultrasonography (EUS) in each stage of Sakita's endoscopic classification. In the second study, 16 patients with gastric ulcer scars were examined by EUS in both S1 stage and S2 stage. Ulcer area, ulcer length, thickness of the ulcer base, and diameter of the crater differed significantly in each endoscopic stage. Ulcer area, ulcer length, and thickness of the ulcer base differed significantly even between S1 stage and S2 stage. Mean ulcer area in S1 stage was 96.9 mm2; this shows that the ulcer scar in S1 stage is incomplete with regard to tissue contraction. Although ulcer area in S2 stage had shrunk significantly compared with that in S1 stage, it still remained 53.3 mm2. This implies that the risk of local recurrence remains even in S2 stage. Sequential observation by EUS showed that the healing of the ulcer surface strongly reflected the healing within the ulcers.


[A treatise on medieval toxicology: the "Liber de venenis" by Pietro d'Abano (French translation from the early 15th century)].
Presentation of the manuscript Paris, B.N.F., Fr. 14820 (French translation of the Liber de venenis by Pietro d'Abano completed in 1402) and description of the treatise from the 14th century, which is a sum of the knowledge on poisons as of that time. The critical study of the manuscript, now in the process of publication, sets forth evidence of significant variations between the original text and the translation.


Bradykinin augments insulin-stimulated glucose transport in rat adipocytes via endothelial nitric oxide synthase-mediated inhibition of Jun NH2-terminal kinase.
An increase in bradykinin has been suggested to contribute to the enhanced insulin sensitivity observed in the presence of ACE inhibitors. To investigate a potential direct, nonvascular effect on an insulin target tissue, the effect of bradykinin on glucose uptake and insulin signaling was studied in primary rat adipocytes. Whereas basal glucose uptake was not altered, bradykinin augmented insulin-stimulated glucose uptake twofold, which was blocked by HOE-140, a bradykinin B2 receptor antagonist. The bradykinin effect on glucose uptake was nitric oxide (NO) dependent, mimicked by NO donors and absent in adipocytes from endothelial NO synthase-/- mice. Investigation of insulin signaling revealed that bradykinin enhanced insulin receptor substrate-1 (IRS-1) Tyr phosphorylation, Akt/protein kinase B phosphorylation, and GLUT4 translocation. In contrast, insulin-stimulated extracellular signal-regulated kinase1/2 and Jun NH2-terminal kinase (JNK) activation were decreased in the presence of bradykinin, accompanied by decreased IRS-1 Ser307 phosphorylation. Furthermore, bradykinin did not enhance insulin action in the presence of the JNK inhibitor, SP-600125, or in adipocytes from JNK1-/- mice. These data indicate that bradykinin enhances insulin sensitivity in adipocytes via an NO-dependent pathway that acts by modulating the feedback inhibition of insulin signaling at the level of IRS-1.


First evidence of bone morphogenetic protein 1 expression and activity in sheep ovarian follicles.
Bone morphogenetic protein (BMP) 1 is a vertebrate metalloproteinase of the astacin family. BMP1 plays a key role in regulating the formation of the extracellular matrix (ECM), particularly by processing the C-propeptide of fibrillar procollagens. BMP1 also promotes BMP signaling by releasing BMP signaling molecules from complexes with the BMP-antagonist chordin. As a result of BMP1's dual role in both ECM formation and BMP signaling, we hypothesized that BMP1 could play a role in ovarian physiology. Using the sheep ovary as a model system, we showed that BMP1 was expressed in the ovary throughout early fetal stages to adulthood. Furthermore, in adult ovaries, BMP1 was expressed along with chordin, BMP4, and twisted gastrulation, which together form an extracellular regulatory complex for BMP signaling. Within ovine ovaries, immunohistochemical localization demonstrated that BMP1 was present in granulosa cells at all stages of follicular development, from primordial to large antral follicles, and that the levels of BMP1 were not affected by the final follicle selection mechanism. In cultured granulosa cells, BMP1 expression was not affected by gonadotropins, but BMP4 and activin A had opposing effects on the levels of BMP1 mRNA. BMP1 appeared to be secreted into the follicular fluid of antral follicles, where it is able to exert procollagen C-proteinase and chordinase activities. Interestingly, BMP1 activity in follicular fluid decreased with follicular growth.


Acute Mastoiditis Associated with Pseudomonas Aeruginosa in the Pediatric Population of the Umbria Region, Italy.
Acute mastoiditis (AM) is the most common complication of acute otitis media (AOM) and is one of the most severe acute bacterial diseases in infants and children. In some geographic areas, the incidence of AM is increasing, and the causative role of some bacterial pathogens could be greater than previously thought. In this paper, the results of a study that evaluated the epidemiology and microbial etiology of paediatric AM in Umbria, which is a region of central Italy, are reported. This is a retrospective study of patients aged 0-14 years with AM admitted to the pediatric wards of the hospitals of Umbria, Italy, between June 1 and September 30 in four consecutive years (2015-2018). A total of 108 children were enrolled. The prevalence of AM in males during the four years of analysis was significantly higher than that in females at 63% (95% confidence intervals [CI]: 0.54-0.72). The most frequently affected age groups were 5-9 years (45.4%) and 10-14 years (31.5%), with statistically significant differences in comparison with children aged <1 year (5.6%, 95% CI: 0.01-0.10) and 1-4 years (17.6%, 95% CI: 0.10-0.25). In most cases (64, 59.3%), AM was associated with spontaneous tympanic membrane perforation (STP). The culture of the middle ear fluid revealed the presence of Pseudomonas aeruginosa in 56 cases (51.6%). The mean incidence rates of pediatric AM in Umbria during the study increased significantly with time, as it was 18.18/100,000 children/year in 2015-2016 and 29.24/100,000 children/year in 2017-2018 (CI difference: +2.5 - +19.9, p < 0.05). The incidence rates of Pseudomonas aeruginosa detection in pediatric AM associated with STP significantly increased with time. The incidence was 6.06/100,000 children/year in 2015-2016 and 18.61/100,000 children/year in 2017-2018 (CI difference: +6.1 - +19.0, p < 0.001). This study demonstrated the high and increasing incidence of AM in the Umbria region during the summer months and the frequent detection of P. aeruginosa as an etiologic agent of the disease in the presence of STP. Confirmation of these results with a larger study population, in different settings, and throughout the whole year is needed to define the first-line approach of AM with STP in pediatrics.


Kerr-nonlinearity induced bistable-like parity-time phase transition in coupled waveguides.
We analyze the parity-time (PT) symmetric phase in coupled two waveguides with a Kerr-type medium in between. Paying attention to the emitted field from a dipole source inside, we show that when the strength of the dipole increases, the optical Kerr effect can render a phase transition from the exact PT phase to the broken PT phase. Furthermore, a salient phenomenon of bistable-like PT phase is observed, in which the emitted field possesses a paradox between the two kinds of PT phases. We show that the physical mechanism of this bistable-like phenomenon is a globally inhomogeneous PT phase, in which different spatial regions of the whole structure can possess different PT phases (broken or exact). This study highlights the potential to manipulate the PT phase transition by using optical nonlinearity for many interesting applications.


Hydration Status of Patients Dialyzed with Biocompatible Peritoneal Dialysis Fluids.
♦ Biocompatible fluids for peritoneal dialysis (PD) have been introduced to improve dialysis and patient outcome in end-stage renal disease. However, their impact on hydration status (HS), residual renal function (RRF), and dialysis adequacy has been a matter of debate. The aim of the study was to evaluate the influence of a biocompatible dialysis fluid on the HS of prevalent PD patients. ♦ The study population consisted of 18 prevalent PD subjects, treated with standard dialysis fluids. At baseline, 9 patients were switched to a biocompatible solution, low in glucose degradation products (GDPs) (Balance; Fresenius Medical Care, Bad Homburg, Germany). Hydration status was assessed through clinical evaluation, laboratory parameters, echocardiography, and bioimpedance spectroscopy over a 24-month observation period. ♦ During the study period, urine volume decreased similarly in both groups. At the end of the evaluation, there were also no differences in clinical (body weight, edema, blood pressure), laboratory (N-terminal pro-brain natriuretic peptide, NTproBNP), or echocardiography determinants of HS. However, dialysis ultrafiltration decreased in the low-GDP group and, at the end of the study, equaled 929 ± 404 mL, compared with 1,317 ± 363 mL in the standard-fluid subjects (p = 0.06). Hydration status assessed by bioimpedance spectroscopy was +3.64 ± 2.08 L in the low-GDP patients and +1.47 ± 1.61 L in the controls (p = 0.03). ♦ The use of a low-GDP biocompatible dialysis fluid was associated with a tendency to overhydration, probably due to diminished ultrafiltration in prevalent PD patients.


Comparison of Survival After In-Hospital Cardiac Arrest in Patients With Versus Without Diabetes Mellitus.
Diabetes mellitus (DM) increases the risk of sudden cardiac death, but the extent to which it influences survival after an in-hospital cardiac arrest (IHCA) remains unclear. We assessed the association of DM and survival after IHCA. The study included 1,009,073 patients aged ≥18 years who underwent cardiopulmonary resuscitation for IHCA between January 2003 and December 2013, recorded in the Nationwide Inpatient Sample database. The outcomes were survival to hospital discharge and discharge disposition assessed using multivariable logistic regression accounting for relevant covariates and clustering. Of the patients with IHCA, 30.8% (310,825) had DM and were more likely to be older and to have a higher prevalence of co-morbidities including hypertension, dyslipidemia, chronic kidney disease, and previous cardiovascular disease (all p <0.001). The rates of survival to hospital discharge after IHCA were 27.0% and 25.1% in patients with and without DM, respectively. After multivariable adjustment, DM was associated with a modestly lower risk-adjusted survival to hospital discharge (adjusted OR [aOR] 0.96, 95% confidence interval [95% CI] 0.95 to 0.97, p <0.001). This association was influenced by a number of factors (all interaction p <0.001), including a lower risk of survival among patients with DM who were younger (aOR 0.93, 95% CI 0.92 to 0.94 among those aged <75 years), those with a primary cardiovascular diagnosis (aOR 0.88, 95% CI 0.86 to 0.89), and those with ventricular fibrillation/ventricular tachycardia as the cardiac arrest rhythm (aOR 0.88, 95% CI 0.79 to 0.82). Patients with DM had lower odds of being discharged home with self-care after surviving an IHCA (p <0.001). In conclusion, preexisting DM was associated with a modestly lower risk-adjusted chance of survival after an IHCA.


Dynamically tunable chemiluminescence of luminol-functionalized silver nanoparticles and its application to protein sensing arrays.
It is still a great challenge to develop an array-based sensing system that can obtain only multiparameters, according to a single experiment and device. The role of conventional chemiluminescence (CL) in biosensing has been limited to a signal transducer in which a single signal (CL intensity) can be obtained for quantifying the concentrations of analytes. In this work, we have developed an dynamically tunable CL system, based on the reaction of luminol-functionalized silver nanoparticles (luminol-AgNPs) with H2O2, which could be tunable via adjusting various conditions such as the concentration of H2O2, pH value, and addition of protein. A single experiment operation could obtain multiparameters including CL intensity, the time to appear CL emission and the time to reach CL peak value. The tunable, low-background, and highly reproducible CL system based on luminol-AgNPs is applied, for the first time, as a sensing platform with trichannel properties for protein sensing arrays by principal component analysis. Identification of 35 unknowns demonstrated a success rate of >96%. The developed sensing arrays based on the luminol-AgNPs provide a new way to use nanoparticles-based CL for the fabrication of sensing arrays and hold great promise for biomedical application in the future.


Mixed Leptonic and Hadronic Corrections to the Anomalous Magnetic Moment of the Muon.
Higher-order hadronic corrections to the anomalous magnetic moment of the muon have been evaluated including next-to-next-to-leading-order insertions of hadronic vacuum polarization and next-to-leading-order corrections to hadronic light-by-light scattering. This leaves a set of mixed leptonic and hadronic corrections in the form of double-bubble topologies as the only remaining hadronic effect at O(α^{4}). Here, we estimate these contributions by analyzing the respective cuts of the diagrams, suggesting that the impact is limited to ≲1×10^{-11} and thus negligible at the level of the final precision of the Fermilab g-2 experiment.


Schwann cell-like differentiated adipose stem cells promote neurite outgrowth via secreted exosomes and RNA transfer.
Adipose derived stem cells can be stimulated to produce a growth factor rich secretome which enhances axon regeneration. In this study we investigated the importance of exosomes, extracellular vesicles released by many different cell types, including stem cells and endogenous nervous system Schwann cells (SCs), on neurite outgrowth. Adipose derived stem cells were differentiated towards a Schwann cell-like phenotype (dADSCs) by in vitro stimulation with a mix of factors (basic fibroblast growth factor, platelet derived growth factor-AA, neuregulin-1 and forskolin). Using a precipitation and low-speed centrifugation protocol the extracellular vesicles were isolated from the medium of the stem cells cultures and also from primary SCs. The conditioned media or concentrated vesicles were applied to neurons in vitro and computerised image analysis was used to assess neurite outgrowth. Total RNA was purified from the extracellular vesicles and investigated using qRT-PCR. Application of exosomes derived from SCs significantly enhanced in vitro neurite outgrowth and this was replicated by the exosomes from dADSCs. qRT-PCR demonstrated that the exosomes contained mRNAs and miRNAs known to play a role in nerve regeneration and these molecules were up-regulated by the Schwann cell differentiation protocol. Transfer of fluorescently tagged exosomal RNA to neurons was detected and destruction of the RNA by UV-irradiation significantly reduced the dADSCs exosome effects on neurite outgrowth. In contrast, this process had no significant effect on the SCs-derived exosomes. In summary, this work suggests that stem cell-derived exosomes might be a useful adjunct to other novel therapeutic interventions in nerve repair.


[PrP protein can bind to protein kinase CK2 both in native and recombinant forms in vitro].
To explore the possible molecular interaction between CK2 and PrP, the full length sequences of human CK2alpha and CK2beta genes were amplified with RT-PCR using the mRNA from cell line SH-SY5Y as the template, and then the fusion proteins HIS-CK2alpha and GST-HIS-CK2beta were expressed in E. coli. The interaction between CK2 and PrP was evaluated with immunoprecipitation and pull-down assays. The results demonstrated that recombinant PrP bound specifically with CK2alpha, but not with CK2beta. The native CK2 and PrP in the hamster brains interacted each other, forming protein complexes. The domain responsible for interacting with CK2alpha was located at the C-terminal segment of PrP (residues 90-231). This study proposed reliable experimental data for the molecular interaction between PrP and CK2alpha, both in recombinant and native categories. These results supply scientific clues for further assessing the potential biological significance of the interaction of PrP with CK2 and possible role of CK2 in the pathogenesis of prion diseases.


Modelling the influence of land-use changes on biophysical and biochemical interactions at regional and global scales.
Land-use changes since the start of the industrial era account for nearly one-third of the cumulative anthropogenic CO2 emissions. In addition to the greenhouse effect of CO2 emissions, changes in land use also affect climate via changes in surface physical properties such as albedo, evapotranspiration and roughness length. Recent modelling studies suggest that these biophysical components may be comparable with biochemical effects. In regard to climate change, the effects of these two distinct processes may counterbalance one another both regionally and, possibly, globally. In this article, through hypothetical large-scale deforestation simulations using a global climate model, we contrast the implications of afforestation on ameliorating or enhancing anthropogenic contributions from previously converted (agricultural) land surfaces. Based on our review of past studies on this subject, we conclude that the sum of both biophysical and biochemical effects should be assessed when large-scale afforestation is used for countering global warming, and the net effect on global mean temperature change depends on the location of deforestation/afforestation. Further, although biochemical effects trigger global climate change, biophysical effects often cause strong local and regional climate change. The implication of the biophysical effects for adaptation and mitigation of climate change in agriculture and agroforestry sectors is discussed.


Relationship Between Cancer Related Fatigue, Physical Activity Related Health Competence, and Leisure Time Physical Activity in Cancer Patients and Survivors.
Background: A large body of evidence supports the positive effects of leisure time physical activity (LTPA) and exercise on cancer survivors. However, only a fraction of survivors manages to attain international PA recommendations. This can be attributed to several external and internal barriers toward PA those patients seem to encounter, with cancer related fatigue (CRF) being the most reported internal barrier. Nevertheless, self-efficacy and knowledge about the utilization of LTPA can serve as facilitators of PA, which also correspond to certain constituents of physical activity related health competence (PAHCO). Since PAHCO is not investigated in cancer survivors we investigated if PAHCO can mediate the negative relationship between CRF and LTPA. Methods: We surveyed 398 cancer survivors with different cancer types and therapy status. The patients completed the EORTC QLQ-FA12 (EORTC FA12) to assess CRF, the PAHCO questionnaire to assess PAHCO and the SQUASH to assess LTPA. We followed a two-step structural equation modeling approach. The first step established the measurement model, in the second step we fitted the mediation model. Since 163 patients chose not to answer the SQUASH, the mediation model was only fitted to the data of the remaining 235 participants. Results: The proposed measurement model of the PAHCO offered an excellent fit. We found small to moderate positive associations between the PAHCO dimensions and the LTPA, and negative moderate relationships between the PAHCO and the EORTC FA12 dimensions. We did not observe a relationship between the EORTC FA12 dimensions and the LTPA (p > 0.05). The hypothesized mediation models did not display an appropriate fit. Conclusion: The PAHCO confirmed its factorial validity; furthermore, it appears to have a positive relationship to LTPA. Therefore, integrating psycho-educational aspects can be beneficial in order to increase the PAHCO in exercise interventions. Because of the cross-sectional character of this study, causal conclusions are not suitable, therefore the longitudinal relationships of LTPA, CRF, and PAHCO require further investigation.


Stimulation of luteinizing hormone release and cyclic nucleotide production by arachidonic acid in cultured pituitary gonadotrophs.
Gonadotropin-releasing hormone (GnRH) stimulates luteinizing hormone (LH) release and cyclic guanosine 3',5-cyclic monophosphate (cGMP) production in rat anterior pituitary cells through a calcium-dependent activation mechanism that involves increased phospholipid turnover and liberation of arachidonic acid. In enriched pituitary gonadotrophs, LH release was stimulated by arachidonic acid and its oxygenated metabolite, 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE), in a dose-dependent manner. The prominent LH responses of purified gonadotrophs to arachidonic acid suggest that the secretory actions of arachidonate are exerted primarily on the gonadotroph and do not involve the participation of other pituitary cell types. Preincubation of pituitary cells with stimulatory concentrations of arachidonic acid for up to 120 min did not alter the subsequent LH responses elicited by GnRH, indicating that the secretory mechanism was unimpaired by arachidonate treatment and that no cross-desensitization occurs during sequential exposure of gonadotrophs to the two stimuli of LH release. Cyclic adenosine 3',5-monophosphate (cAMP) production was stimulated by 10 microM arachidonic acid to the same degree (about 2-fold) as by GnRH, but did not parallel the progressive LH response to higher arachidonate concentrations. cGMP production was initially stimulated by addition of arachidonic acid but returned to the control value after 5 min, whereas GnRH typically elicited a prolonged cGMP response. In contrast to the calcium-independent action of arachidonic acid, the stimulatory effect of 5-HETE on LH release required the presence of extracellular Ca2+, as previously observed for GnRH. These findings demonstrate that arachidonic acid and its metabolite, 5-HETE, partially reproduce the actions of GnRH upon LH release and cyclic nucleotide production.(ABSTRACT TRUNCATED AT 250 WORDS)


Differential Behavioral and Neural Effects of Regional Cerebellar tDCS.
The human cerebellum contributes to both motor and non-motor processes. Within the cerebellum, different subregions support sensorimotor and broader cognitive functions, due to regional patterns in anatomical connectivity with the cerebral cortex and spinal and vestibular systems. We evaluated the effects of transcranial direct current stimulation (tDCS) targeting different cerebellar regions on language task performance and whole-brain functional activation patterns. Functional MRI data were acquired while 43 healthy young adults (15 males, 28 females; 23.3 ± 3.0 years) performed a sentence completion task before and after 20 min of 1.5 mA anodal tDCS. Participants received tDCS targeting either the anterior sensorimotor cerebellum (n = 11; 3 cm right of inion, over lobule V); the right posterolateral cerebellum (n = 18; 1 cm down and 4 cm right of inion, over lobule VII); or sham tDCS (n = 14). TDCS targeting the right posterolateral cerebellum improved task accuracy relative to the sham condition (p = 0.04) and increased activation in left frontal and temporal cortices relevant to task performance (post-tDCS > pre-tDCS; T 3.17, FDR p < 0.05 cluster correction). The regions of increased BOLD signal after right posterolateral cerebellar tDCS fell within the network showing functional connectivity with right cerebellar lobule VII, suggesting specific modulation of this network. In contrast, tDCS targeting the sensorimotor cerebellum did not impact task performance and increased BOLD signal only in one cluster extending into the precentral gyrus. These findings indicate that sensorimotor and cognitive functional cerebellar subregions differentially impact behavioral task performance and task-relevant activation patterns, further contributing to our understanding of the cerebellar modulation of motor and non-motor functions.


Magnetic nanoparticle-antibody conjugates for the separation of Escherichia coli O157:H7 in ground beef.
The immunomagnetic separation with magnetic nanoparticle-antibody conjugates (MNCs) was investigated and evaluated for the detection of Escherichia coli O157:H7 in ground beef samples. MNCs were prepared by immobilizing biotin-labeled polyclonal goat anti-E. coli antibodies onto streptavidin-coated magnetic nanoparticles. For bacterial separation, MNCs were mixed with inoculated ground beef samples, then nanoparticle-antibody-E. coli O157:H7 complexes were separated from food matrix with a magnet, washed, and surface plated for microbial enumeration. The capture efficiency was determined by plating cells bound to nanoparticles and unbound cells in the supernatant onto sorbitol MacConkey agar. Key parameters, including the amount of nanoparticles and immunoreaction time, were optimized with different concentrations of E. coli O157:H7 in phosphate-buffered saline. MNCs presented a minimum capture efficiency of 94% for E. coli O157:H7 ranging from 1.6 x 10(1) to 7.2 x 10(7) CFU/ml with an immunoreaction time of 15 min without any enrichment. Capture of E. coli O157:H7 by MNCs did not interfere with other bacteria, including Salmonella enteritidis, Citrobacter freundii, and Listeria monocytogenes. The capture efficiency values of MNCs increased from 69 to 94.5% as E. coli O157:H7 decreased from 3.4 x 10(7) to 8.0 x 10(0) CFU/ml in the ground beef samples prepared with minimal steps (without filtration and centrifugation). An enrichment of 6 h was done for 8.0 x 10(0) and 8.0 x 10(1) CFU/ml of E. coli O157:H7 in ground beef to increase the number of cells in the sample to a detectable level. The results also indicated that capture efficiencies of MNCs for E. coli O157:H7 with and without mechanical mixing during immunoreaction were not significantly different (P > 0.05). Compared with microbeads based immunomagnetic separation, the magnetic nanoparticles showed their advantages in terms of higher capture efficiency, no need for mechanical mixing, and minimal sample preparation.


The many facets of unawareness in huntington disease.
Unawareness or diminished awareness is present when a patient's perception of obvious disease manifestations and impact differ from that of observers such as clinicians or family members. We examined studies that specifically investigate unawareness in Huntington disease (HD). Unawareness of motor, cognitive, behavioral, and functional aspects of HD has been documented throughout the disease course. This can occur at motor and cognitive onset but is more pronounced as the disease progresses. We discuss the implications for diagnosis, symptom report at presentation, timing of diagnosis, acceptance of symptomatic care strategies, and reporting in clinical trials. Assessments of work place competency, discrimination, driving, and the particular challenges of isolated patients without caregivers are described. Engaging with a person who is unaware of their disease or its impact presents a number of conflicts, including maintaining the right to autonomy, privacy, confidentiality, and independence while recognizing concerns for the wellbeing of the vulnerable person with HD and their caregiver when the unaware person refuses assistance. Unawareness is seen increasingly as neurologically based due to the impairment of functional networks, predominantly in nondominant frontostriatal pathways.


Implementation and findings on a one-minute sit-stand test for prehospital triage in patients with suspected COVID-19-a pilot project.
During the initial Coronavirus Disease 2019 (COVID-19) pandemic wave, sparse personal protection equipment made telephone triage of suscpeted COVID-19 patients for ambulance transport necessary. To spare resources, stable patients were often treated and released on-scene, but reports from Italy suggested that some later detoriated. We implemented a prehospital sit-stand test to identify patients in risk for detoriation. The test was implemented as part of a new guideline in stable suspected COVID-19 patients younger than 70 years with no risk factors for serious disease triaged by general practitioners to ambulance response in the Central Denmark Region. Data were collected from April 6th to July 6th 2020. The primary outcome for this study was the proportion of patients treated with oxygen within 7 days among patients decompensating vs patients not decompensating during the test. Data on 156 patients triaged to ambulance response by general practioners were analysed. In total 86/156 (55%) were tested with the sit-stand test. Due to off-guideline use of the test, 30/86 (34.8%) were either older than 70 or had risk factors for serious disease. 10/156 (6%) of patients had a positive COVID-19-test. In total, 17/86 (20%) decompensated during the test and of these, 9/17 (53%) were treated with oxygen compared to 2/69 (3%) in patients who did not decompensate (p < 0.001). In a population suspected of COVID-19 but with a low COVD-19 prevalence, decompensation with the sit-stand test was observed in 20% of patients and was associated with oxygen treatment within 7 days. These findings are hypotheses-generating and suggest that physical exercise testing may be usefull for decision making in emergency settings.


A new approach for reducing beam hardening artifacts in polychromatic X-ray computed tomography using more accurate prior image.
Metal artifacts severely degrade CT image quality in clinical diagnosis, which are difficult to removed, especially for the beam hardening artifacts. The metal artifact reduction (MAR) based on prior images are the most frequently-used methods. However, there exists a lot misclassification in most prior images caused by absence of prior information such as the spectrum distribution of X-ray beam source, especially many or big metal included. The purpose of this work is to find a more accurate prior image to improve image quality. The proposed method comprise of following four steps. First, the metal image is segmented by thresholding an initial image, where the metal traces are identified in the initial projection data using the forward projection of the metal image. Second, the accurate absorbent model of certain metal image is calculated according to the spectrum distribution of certain X-ray beam source and energy-dependent attenuation coefficients of metal. Then, a new metal image is reconstructed by the general analytical reconstruction algorithm such as filtered back projection (FPB). The prior image is obtained by segmenting the difference image between the initial image and the new metal image into air, tissue and bone. Finally, the initial projection data are normalized by dividing the projection data of prior image pixel to pixel, the corrected image is obtained by interpolation, denormalization and reconstruction. Some clinical images with dental fillings and knee prostheses are used to evaluate the proposed algorithm and normalized metal artifact reduction (NMAR) and linear interpolation (LI) method. The results demonstrate the artifacts can be reduced efficiently by the proposed method. The proposed method could obtain an exact prior image using the prior information about X-ray beam source and energy-dependent attenuation coefficients of metal. As a result, the better performance of reducing beam hardening artifacts can be improved, even though there were many or big implants. Moreover, the process of the proposed method is rather simple and little extra calculation burden is necessary. It has superiorities over other algorithms when include big or many implants.


How well do nurses recognize malnutrition in elderly patients?
Malnutrition is a common and underrecognized clinical problem among aged institutionalized patients. The aim of this study was to investigate how well nurses recognize malnutrition in elderly patients in long-term care hospitals in Helsinki. In this descriptive, cross-sectional study, the nutritional status of 1043 elderly patients was assessed with the Mini Nutritional Assessment (MNA), their body mass indices (BMIs) (kg m(-2)) were counted, and factors related to their nutritional care were queried using a structured questionnaire. In addition, we asked the opinions of 53 nurses on whether they considered their patients to suffer from malnutrition. All the long-term care hospitals in Helsinki, Finland participated in this study. The mean age of the patients was 81 years. The nurses considered only 15.2% of the patients to be malnourished, although the MNA showed that 56.7% were malnourished (MNA<17 points). Those recognized as malnourished were truly anorectic, with a mean BMI of 17.2. Of those patients having a BMI<20 and MNA<17, the nurses considered only one-third to be malnourished. Of those having a BMI>24 but MNA<17, only 2% were recognized as having malnutrition. Even those patients considered to be malnourished received snacks and nutritional supplements less than the patients that the nurses considered to have normal nutritional status. However, only one in six of the malnourished patients received oral nutritional supplements. The nurses recognized malnutrition in their aged patients poorly. Nutrition education for nurses is urgently needed, as malnutrition and weight loss have been considered significant problems, and the benefits of nutritional care are well established.


The effect of smoking on early chorionic villous vascularisation.
The aim of the study was to investigate whether first trimester chorionic villous vascularisation is different in women who smoked cigarettes before and during pregnancy in comparison with women who did not smoke. Placentas of smoking (>10 cigarettes/day, n = 13) and non-smoking women (n = 13), scheduled for a legal termination of a viable first trimester pregnancy for social indications, were retrieved. Placental tissues of 3-5 mm³ were whole mount CD31 immunofluorescence stained. Images of the CD31 immunofluorescence and contour of the villi were captured using an Optical Projection Tomography scanner. An immersive BARCO virtual reality system was used to create an enlarged interactive 3-dimensional hologram of the reconstructed images. Automatic volume measurements were performed using a flexible and robust segmentation algorithm that is based on a region-growing approach in combination with a neighbourhood variation threshold. The villous volume, vascular volume and vascular density were measured for the total chorionic villous tree as well as for its central and peripheral parts. No differences in maternal age and gestational age were found between non-smoking and smoking women. No differences were found in the total, central and peripheral villous tree volume and vascular volume. The central (13.4% vs. 9.5%, p=0.03) and peripheral (8.4% vs. 6.4%, p=0.02) villous tree vascular densities were increased in the smoking women as compared with the non-smoking women. In conclusion, chorionic villous vascularisation is already altered in first trimester of pregnancy in women who smoked cigarettes before and during pregnancy.


Novel ASK1 Inhibitor AGI-1067 Attenuates AGE-Induced Fibrotic Response by Suppressing the MKKs/p38 MAPK Pathway in Human Coronary Arterial Smooth Muscle Cells.
The phenotype shifting of vascular smooth muscle cells (VSMCs) was indicated to play a role during the initial stage of atherosclerotic plaque formation by facilitating extracellular matrix deposition. This study was aimed at investigating the involvement of the apoptosis signal-regulating kinase 1 (ASK1) /mitogen-activated protein kinase (MAPK) kinases (MKKs) /p38 MAPK pathway in the advanced glycation end product (AGE) -induced fibrotic response of VSMCs. The effect of the novel ASK1 inhibitor AGI-1067 was also studied.Cultured human coronary smooth muscle cells (HCSMCs) were exposed to AGEs. AGI-1067 and siRNAs silencing mkk3, mkk6, and p38 mapk were used to treat the cells. The activation of MKK3, MKK6, and p38 MAPK was assessed by immunoblotting. Fibrotic response was assessed by the fluorescence immunohistochemistry staining of collagen I and collagen VIII. Activation of immunoprecipitation determined the association of ASK1 and its inhibitor thioredoxin. A kinase assay was used to determine ASK1 activity.AGE incubation significantly activated ASK1, MKK3, and MKK6, which led to activation of p38 MAPK, resulting in upregulated fibrotic response in HCSMCs. However, siRNAs knocking down mkk3, mkk6, and p38 mapk impaired this fibrotic response. AGI-1067 administration not only dramatically inhibited the activation of ASK1/MKKs/p38 MAPK but also suppressed the expression of the downstream proteins, including transforming growth factor-β1, connective tissue growth factor, collagen I, and collagen VIII in HCSMCs exposed to AGEs.The ASK1/MKKs/p38 MAPK pathway was activated by AGEs, leading to the fibrotic response in VSMCs. AGI-1067 reversed this process by maintaining the inactive state of ASK1.


Frequent abnormalities of the putative tumor suppressor gene FHIT at 3p14.2 in pancreatic carcinoma cell lines.
The FHIT gene is localized on chromosome 3p14, a region including a tumor cell-specific, commonly deleted region. To determine the role of the FHIT gene in pancreatic carcinogenesis, 14 pancreatic carcinoma cell lines were analyzed by reverse transcription-PCR and exon-specific PCR amplification of genomic DNA. The full-length FHIT transcript was lost in 70% of the pancreatic carcinoma cell lines analyzed, while 66% also revealed intragenic homozygous deletions of exons 3, 4, and 5. Truncated FHIT transcripts lacking a variable number of exons most likely represented alternative splicing products. Fhit protein expression was dependent on a full-length FHIT transcript. The results suggest that the FHIT gene may be a target tumor suppressor gene involved in pancreatic carcinogenesis.


SRC-family kinases are activated in non-small cell lung cancer and promote the survival of epidermal growth factor receptor-dependent cell lines.
The role of Src-family kinases (SFKs) in non-small cell lung cancer (NSCLC) has not been fully defined. Here we addressed this question by examining SFK phosphorylation in NSCLC biopsy samples and using genetic and pharmacological approaches to inhibit SFK expression and activity in cultured NSCLC cells. Immunohistochemical analysis of NSCLC biopsy samples using a Tyr416 phosphorylation-specific, pan-SFK antibody revealed staining in 123 (33%) of 370 tumors. Because c-Src is known to be both an upstream activator and downstream mediator of epidermal growth factor receptor (EGFR), we next investigated SFK phosphorylation in a panel of NSCLC cell lines, including ones that depend on EGFR for survival. The EGFR-dependent NSCLC cell lines HCC827 and H3255 had increased phosphorylation of SFKs, and treatment of these cells with an SFK inhibitor (PP1 or SKI-606) induced apoptosis. PP1 decreased phosphorylation of EGFR, ErbB2, and ErbB3 and strikingly enhanced apoptosis by gefitinib, an EGFR inhibitor. HCC827 cells transfected with c-Src short hairpin RNA exhibited diminished phosphorylation of EGFR and ErbB2 and decreased sensitivity to apoptosis by PP1 or gefitinib. We conclude that SFKs are activated in NSCLC biopsy samples, promote the survival of EGFR-dependent NSCLC cells, and should be investigated as therapeutic targets in NSCLC patients.


Terrestrial species adapted to sea dispersal: Differences in propagule dispersal of two Caribbean mangroves.
A central goal of comparative phylogeography is to understand how species-specific traits interact with geomorphological history to govern the geographic distribution of genetic variation within species. One key biotic trait with an immense impact on the spatial patterns of intraspecific genetic differentiation is dispersal. Here, we quantify how species-specific traits directly related to dispersal affect genetic variation in terrestrial organisms with adaptations for dispersal by sea, not land-the mangroves of the Caribbean. We investigate the phylogeography of white mangroves (Laguncularia racemosa, Combretaceae) and red mangroves (Rhizophora mangle, Rhizophoraceae) using chloroplast genomes and nuclear markers (thousands of RAD-Seq loci) from individuals throughout the Caribbean. Both coastal tree species have viviparous propagules that can float in salt water for months, meaning they are capable of dispersing long distances. Spatially explicit tests of the role of ocean currents on patterning genetic diversity revealed that ocean currents act as a mechanism for facilitating dispersal, but other means of moving genetic material are also important. We measured pollen- vs. propagule-mediated gene flow and discovered that in white mangroves, seeds were more important for promoting genetic connectivity between populations, but in red mangroves, the opposite was true: pollen contributed more. This result challenges our concept of the importance of both proximity to ocean currents for moving mangrove seeds and the extent of long-distance pollen dispersal. This study also highlights the importance of spatially explicit quantification of both abiotic (ocean currents) and biotic (dispersal) factors contributing to gene flow to understand fully the phylogeographic histories of species.


[Hindfoot arthrodesis resulting from retrograde medullary pinning].
Simultaneous arthrodesis of the ankle and subtalar joints and correction of axial malalignment of the hindfoot in cases of bony defects and/or circulatory disturbances of the talus after multiple previous interventions. Internal stabilization with a short distal femur nail. Restoration of pain-free weight bearing. Failure of arthrodesis of the ankle and subtalar joint in patients with severely altered bone structure particularly at the level of the talar dome. Malalignment of hind- and/or forefoot after previous arthrodesis of the ankle and subtalar joint. Poor skin or soft-tissue condition. Acute osteitis/osteomyelitis. Posterolateral approach. Resection of the articular cartilage and the areas of sclerosis of the ankle and posterior facet of the subtalar joint. Interposition of bone grafts harvested from the posterior iliac crest. Correction of malalignment of the hind- and forefoot. Locked nailing with a short distal femur nail. All 21 prospectively enrolled patients were followed-up clinically and radiographically at an average of 1.2 years (0.6-2.1 years) postoperatively. The average age of the 4 women and 17 men at the time of surgery was 53.4 years (38.9-73.7 years). The goal of the surgery was achieved in all patients. Subjective assessment was good in 14 patients and satisfactory in 3 patients. Complications occurred in 5 patients; these included loss of nail purchase (2), dislocation of locking screw (1), breakage of locking screw (1), and nonunion (1).


[A prediction study on the supply of and demand for pharmacists in Japan].
Recently, the separation between dispensing and prescribing drugs has made rapid progress and the number of pharmacists' jobs at hospitals have increased, which means that pharmacists are contributing more than ever to medical treatment. Thus the relation between the supply of and demand for pharmacists has become a topic in the improvement of pharmacist quality. In this study, we used date on pharmacists including type of job, registration number, etc. to predict the relation between the supply of pharmacists at work and the demand for pharmacists. In addition, we analyzed the working trends of pharmacy pharmacists and predicted the in relation between the supply and demand. Due to progress in the separation of dispensing and prescribing drugs, the supply of pharmacists will nearly equal demand temporarily. However, the supply at work will always exceed demand, and we concluded that no shortage of pharmacists would occur. In addition, after the demand for pharmacists stops increasing, the excess supply of pharmacists will continue to increase. Based on analysis of working trends of pharmacy pharmacists, the in number will increase at the same rate as between 1996 and 1998 (approximately 6000 persons/year). Under this assumption, we concluded that no short-age of pharmacy pharmacists will occur, even if the separation of dispensing and prescribing drugs continues to occur at the rate of 5%/year.


Effects of Arousal on Mouse Sensory Cortex Depend on Modality.
Changes in arousal modulate the activity of mouse sensory cortex, but studies in different mice and different sensory areas disagree on whether this modulation enhances or suppresses activity. We measured this modulation simultaneously in multiple cortical areas by imaging mice expressing voltage-sensitive fluorescent proteins (VSFP). VSFP imaging estimates local membrane potential across large portions of cortex. We used temporal filters to predict local potential from running speed or from pupil dilation, two measures of arousal. The filters provided good fits and revealed that the effects of arousal depend on modality. In the primary visual cortex (V1) and auditory cortex (Au), arousal caused depolarization followed by hyperpolarization. In the barrel cortex (S1b) and a secondary visual area (LM), it caused only hyperpolarization. In all areas, nonetheless, arousal reduced the phasic responses to trains of sensory stimuli. These results demonstrate diverse effects of arousal across sensory cortex but similar effects on sensory responses.


Risk assessment of mechanism-based inactivation in drug-drug interactions.
Drug-drug interactions (DDIs) that occur via mechanism-based inactivation of cytochrome P450 are of serious concern. Although several predictive models have been published, early risk assessment of MBIs is still challenging. For reversible inhibitors, the DDI risk categorization using [I]/K(i) ([I], the inhibitor concentration; K(i), the inhibition constant) is widely used in drug discovery and development. Although a simple and reliable methodology such as [I]/K(i) categorization for reversible inhibitors would be useful for mechanism-based inhibitors (MBIs), comprehensive analysis of an analogous measure reflecting in vitro potency for inactivation has not been reported. The aim of this study was to evaluate whether the term λ/k(deg) (λ, first-order inactivation rate at a given MBI concentration; k(deg), enzyme degradation rate constant) would be useful in the prediction of the in vivo DDI risk of MBIs. Twenty-one MBIs with both in vivo area under the curve (AUC) change of marker substrates and in vitro inactivation parameters were identified in the literature and analyzed. The results of this analysis show that in vivo DDIs with >2-fold change of object drug AUC can be identified with the cutoff value of λ/k(deg) = 1, where unbound steady-state C(max) is used for inhibitor concentration. However, the use of total C(max) led to great overprediction of DDI risk. The risk assessment using λ/k(deg) coupled with unbound C(max) can be useful for the DDI risk evaluation of MBIs in drug discovery and development.


Integration of hepatic lipidomics and transcriptomics reveals the effect of butter-derived ruminant trans fatty acids on lipid metabolism in C57BL/6J mice.
Dysregulation of lipid metabolism results in metabolism-related diseases. Our previous research indicated that 1.3% E and 4% E ruminant trans fatty acids (R-TFA) caused dyslipidemia and promoted atherosclerotic plaques in ApoE-/- mice, presenting detrimental effects. However, the effect of R-TFA on the lipid metabolism of normal mice remains unclear. Therefore, our current research aims to explore the effects of butter-derived R-TFAs on the lipid metabolism of C57BL/6J mice through the integration of lipidomics and transcriptomics. As a result, we found that 1.3% E butter-derived R-TFA promoted dyslipidemia and impaired hepatic function in C57BL/6J mice fed a high-fat diet, which was associated with an increase in DG (18:1/22:5), TG (18:1/18:2/22:4) and FA (24:5) as determined through lipidomics analysis, but had a less significant effect on C57BL/6J mice fed a low-fat diet. Through a combination analysis and verification of gene expression, we found that the arachidonic acid pathway might be involved in the disruption of lipid metabolism by butter-derived R-TFA. In addition, butter-derived R-TFA up-regulated the expression of unigene thromboxane-A synthase 1 (Tbxas1), arachidonate lipoxygenase 3 (Aloxe3), acyl-coenzyme A thioesterase 2 (Acot2), epoxide hydrolase 2 (Ephx2) and carbonyl reductase 3 (Cbr3) in C57BL/6J mice fed a high-fat diet. Herein, our research provides a new perspective for exploring the effects of butter-derived R-TFA on lipid metabolism and speculates on the possible mechanism of lipid metabolism disorder induced by butter-derived R-TFA in C57BL/6J mice fed a high-fat diet.


Mycetoma in Uganda: A neglected tropical disease.
Mycetoma is considered a neglected tropical disease globally. However, data on its burden and the associated complications in Uganda are limited. Hence we aimed to estimate its burden in Uganda. Firstly, a systematic PubMed search for all studies of any design on mycetoma in Uganda without restriction to the year of publication was conducted. A retrospective review of all the biopsy reports at the Pathology Reference Laboratory, Department of Pathology, Makerere University, Kampala, Uganda from January 1950 to September 2019 was conducted to identify any reports on mycetoma histological diagnosis. During the 70-years study period, 30 cases were identified by the literature review, with 249 additional cases identified by review of biopsy reports (total of 279 cases). The average incidence was estimated at 0.32/100,000 persons and prevalence of 8.32/100,000 persons per decade. However, there was a general decline in the number of cases detected recently. Males and the age group of 21-30 years were the most affected by mycetoma in Uganda, and only 7% of the cases were children. The highest number of cases was recorded from Kampala (n = 30) and Jinja (n = 19) districts. The majority of the cases (68%) were referred from surgical units. The foot was the most affected part of the body (72%). Ten per cent of the cases had bone involvement of which 58% required amputation. Fungi were the most common causative agents (89%) followed by Nocardia species (5%) and Actinomycetes (4%). The index of clinical suspicion of mycetoma was low (45%) with a very large differential diagnosis. Mycetoma is a relatively rare disease in Uganda, mostly caused by fungi, and there is a big gap in data and epidemiological studies. More systematic studies are warranted to define the true burden of mycetoma in Uganda.


The protection of the myocardium by amifostine against mitoxantrone-induced acute cardiotoxicity in rats.
Amifostine (AMI) has been used for the prevention of doxorubicin-induced cardiotoxicity in several experimental and a few clinical studies. The aim of this study was to investigate the effects of AMI on lipid peroxidation, protective enzymes, and mitoxantrone (MITO)-induced acute cardiotoxicity in the rat heart using biochemical tests and histopathological examinations. Thirty-six rats were divided into six groups (n=6 in each). Control rats were given intraperitoneal (i.p.) serum saline and AMI group rats were given 200 mg/kg AMI i.p. Rats received MITO-2.5 and 5 mg/kg i.p. in the MITO-2.5 and MITO-5 groups. AMI 200 mg/kg i.p. was administered 30 min. before the same doses of MITO in the MITO-2.5+AMI and MITO-5+AMI groups. The levels of cardiac enzymes such as creatinine phosphokinase-myocardial band and cardiac troponin T did not change. Malondialdehyde (MDA) levels increased in MITO groups compared to controls. Catalase and glutathione (GSH) levels in the MITO and MITO+AMI groups were higher than in controls. Superoxide dismutase and glutathione peroxidase levels were not different between MITO groups and controls. There was no difference in MDA levels between MITO+AMI groups and controls. Calcium deposition was not detected. The scores of fibrosis, apoptosis, inflammation, and degeneration in MITO groups were higher than in controls. The scores of fibrosis, degeneration and inflammation in MITO+AMI groups were lower. MITO caused lipid peroxidation and myocardial damage, and the myocardium increased catalase and GSH levels to prevent this damage. AMI can protect against MITO-induced acute cardiotoxicity, decreasing myocardial damage and lipid peroxidation.


Study of early events in the protein folding of villin headpiece using molecular dynamics simulation.
Protein folding is scientifically and computationally challenging problem. The early phases of protein folding are interesting due to various events like nascent secondary structure formation, hydrophobic collapse leading to formation of non-native or meta-stable conformations. These events occur within a very short time span of 100 ns as compared to total folding time of few microseconds. It is highly difficult to observe these events experimentally due to very short lifetime. Molecular dynamics simulation technique can efficiently probe the detailed atomic level understanding about these events. In the present paper, all atom molecular dynamics simulation trajectory of nearly 200 ns was carried out for fully solvated villin headpiece with PME treatment using AMBER 7 package. Initial hydrophobic collapse along with secondary structure formation resulted into formation of partially stable non-native conformations. The formation of secondary structural elements and hydrophobic collapse takes place simultaneously in the folding process.


Vanilloid type 1 receptors (VR1) on trigeminal sensory nerve fibres play a minor role in neurogenic dural vasodilatation, and are involved in capsaicin-induced dural dilation.
Capsaicin, the active substance in chilli peppers, activates the vanilloid type 1 receptor (VR1) rather than the vanilloid-like receptor (VRL1) in the trigeminal ganglion and nucleus of small and medium C- and Adelta-fibres. Capsaicin induces calcitonin gene-related peptide (CGRP) release when VR1 receptors are activated, and this can be reversed by both the VR1 receptor antagonist capsazepine and the CGRP blocker alphaCGRP8-37 in vitro. In this study we used intravital microscopy to look at the possible role of the VR1 receptor in the trigeminovascular system in producing dilation of dural blood vessels. Capsazepine (3 mg kg-1) was given to study the effect of the VR1 receptor in dural vessel dilation produced by either electrical stimulation, CGRP (1 microg x kg-1) or capsaicin (7 microg x kg-1) bolus injection. We also looked at the effect of the CGRP blocker alphaCGRP8-37 (300 microg x kg-1) on capsaicin-induced dilation so that we could see if the results found in vitro could also be found in vivo. Electrical stimulation of the dura mater produced a robust vasodilator response between 130 and 137% of baseline diameter that was no different across four repeat stimuli (F3,18=0.6, P=0.61). CGRP similarly produced a dilatation of 99-111% that was no different across four baseline infusions (F3,15=2.4, P=0.113). Capsaicin also produced a consistent dilation of between 112 and 120% of baseline across three injections (F2,10=0.6, P=0.567). Capsazepine did not inhibit the dilation brought about by either electrical stimulation or CGRP injection, while it was able to inhibit the dilation brought about by capsaicin (t5=3.4, P<0.05). AlphaCGRP8-37 also inhibited the capsaicin-induced dilation (t5=7.4, P<0.05) probably inhibiting the action of released CGRP at the CGRP receptor. The study demonstrates that capsaicin can repeatedly induce dural vessel dilation in vivo, presumably through inducing CGRP release from trigeminal sensory nerve fibres, while C-fibres may have been desensitised. The data imply that the VR1 receptor plays only a minor role in trigeminovascular-induced dural vessel dilation.


Ocular pathogenicity of herpes simplex virus.
As a relatively small, discrete organ that contains a number of widely different cell types the eye provides an intriguing system in which to study fundamental aspects of virus/cell interactions. Such aspects are considered with particular reference to herpes simplex virus and the pivotal role of virus/neuron interactions in the development of ocular disease. Three aspects of this interaction are discussed: the entry of virus into the eye latency in the trigeminal ganglion nerve damage.


A high-throughput multiplex method adapted for GMO detection.
A high-throughput multiplex assay for the detection of genetically modified organisms (GMO) was developed on the basis of the existing SNPlex method designed for SNP genotyping. This SNPlex assay allows the simultaneous detection of up to 48 short DNA sequences (approximately 70 bp; "signature sequences") from taxa endogenous reference genes, from GMO constructions, screening targets, construct-specific, and event-specific targets, and finally from donor organisms. This assay avoids certain shortcomings of multiplex PCR-based methods already in widespread use for GMO detection. The assay demonstrated high specificity and sensitivity. The results suggest that this assay is reliable, flexible, and cost- and time-effective for high-throughput GMO detection.


MDMX is a prognostic factor for non-small cell lung cancer and regulates its sensitivity to cisplatin.
Chemoradiotherapy is the standard treatment modality for advanced non-small cell lung cancer (NSCLC). However, drug and radiation resistance remain major factors influencing its clinical outcome. The purpose of this study was to evaluate whether MDMX can affect the chemosensitivity and clinical outcome of NSCLC. Quantitative real-time PCR (qRT-PCR) was performed to assess MDMX mRNA expression levels in 105 primary NSCLC tissues, its corresponding non-cancerous tissues and two NSCLC-derived cell lines (A549 and SK-MES-1). In addition, immunohistochemistry was carried out to detect MDMX protein expression in the primary NSCLC tissues. The MDMX expression levels were correlated with clinicopathological and survival features. The effects of MDMX expression knockdown on NSCLC cell proliferation and chemosensitivity were evaluated using MTT, flow cytometry and soft agar colony assays. We found that the mRNA expression level of MDMX in NSCLC tissues was significantly higher than that in its corresponding non-tumorous tissues. High MDMX expression was found to be related to poor tumor cell differentiation, advanced TNM stages and the occurrence of lymph node metastases. Patients with a high MDMX expression level exhibited a lower overall survival rate than those with a low expression level. Multivariate analysis showed that a high MDMX protein expression level may serve as an independent prognostic factor for NSCLC patients. In addition, we found that MDMX expression knockdown combined with cisplatin treatment in vitro significantly increased apoptosis and decreased soft agar colony formation in NSCLC-derived cells. Our data indicate that MDMX expression may serve as an independent unfavorable prognostic factor for NSCLC patient outcome, which in turn may at least partly be due to the ability of the MDMX protein to regulate the proliferative capacity and chemosensitivity of NSCLC cells.


[Willingness of HIV non-occupational post-exposure prophylaxis and related factors among men who have sex with men in Beijing].
Objective: To examine the willingness of HIV non-occupational post-exposure prophylaxis (nPEP) among MSM and factors related were also assessed. Methods: The respondent-driven sampling method was used to recruit MSM for a face-to-face interview with a structured questionnaire,the sample size was estimated at 600 subjects. Demographic data, sexual behavior, awareness, and willingness regarding nPEP were collected. The factors related to willingness to nPEP were assessed using complex logistic regression. Results: A total of 14 rounds were recruited and 608 MSM subjects were included in the study. The average age was (41.6±11.0) years. 55.4% (95%CI: 49.4%-59.4%) were aware of nPEP, and 4.5% (95%CI: 2.9%-6.2%) have used its. 35.9% (95%CI: 31.1%-40.7%) expressed interest in taking nPEP if needed. Among the reasons for not being willing to take nPEP, 68.9% (244/354) were never heard of nPEP, and 24.6% (87/354) were a fluke mentality. The multivariate logistic analysis results showed that the willingness of nPEP awareness of MSM was related to the group aged 25-39 years old (aOR=1.80, 95%CI: 1.01-3.20), knowing a group of HIV prevention knowledge (aOR=2.43, 95%CI: 1.52-3.90), group of consistent condom use in anal sex in the past half of year (aOR=1.76, 95%CI: 1.11-2.79). Conclusions: The use rate of nPEP among MSM in Beijing was low, and the willingness to use in the future also needs to be improved. The training of social organizations should be strengthened to improve the role of peer education in promoting nPEP.


Volumetric growth rates of untreated cavernous sinus meningiomas.
Meningiomas that arise primarily within the cavernous sinus are often believed to be more indolent in their growth pattern. Despite this perceived growth pattern, disabling symptoms can arise even with small tumors. While research has been done on cavernous sinus meningiomas (CSMs) and their treatment, very little is known about their natural growth rates. With a better understanding of the growth rate of CSM, patient treatment and guidance can be can optimized and individualized. The goal of this study was to determine volumetric growth rates of untreated CSMs. Thirty-seven patients with 166 MR images obtained between May 2004 and September 2019 were reviewed, with a range of 2-13 MR images per patient (average of 4.5 MR images per patient). These scans were obtained over an average follow-up period of 45.9 months (median 33.8, range 2.8-136.9 months). All imaging prior to any intervention was included in this analysis. Volumetric measurements were performed and assessed over time. The estimated volumetric growth rate was 23.3% per year (95% CI 10.2%-38.0%, p < 0.001), which is equivalent to an estimated volume doubling time (VDT) of 3.3 years (95% CI 2.1-7.1 years). There was no significant relationship between growth rate and patient age (p = 0.09) or between growth rate and patient sex (p = 0.78). The median absolute growth rate was 41% with a range of -1% to 1793%. With a definition of "growth" as an increase of greater than 20% during the observed period, 65% of tumors demonstrated growth within their observation interval. Growth rates for each tumor were calculated and tumors were segmented based on growth rate. Of 37 patients, 22% (8) demonstrated no growth (< 5% annual growth, equivalent to a VDT > 13.9 years), 32% (12) were designated as slow growth (annual growth rate 5%-20%, VDT 3.5-13.9 years), 38% (14) were found to have medium growth (annual growth rate 20%-100%, VDT 0.7-3.5 years), and 8% were considered fast growing (annual growth rate > 100%, VDT < 0.7 years). This study evaluated CSM volumetric growth rates. A deeper understanding of the natural history of untreated CSMs allows for better counseling and management of patients.


The ethics of Canadian entry-to-practice pain competencies: how are we doing?
Although unrelieved pain continues to represent a significant problem, prelicensure educational programs tend to include little content related to pain. Standards for professional competence strongly influence curricula and have the potential to ensure that health science students have the knowledge and skill to manage pain in a way that also allows them to meet professional ethical standards. To perform a systematic, comprehensive examination to determine the entry-to-practice competencies related to pain required for Canadian health science and veterinary students, and to examine how the presence and absence of pain competencies relate to key competencies of an ethical nature. Entry-to-practice competency requirements related to pain knowledge, skill and judgment were surveyed from national, provincial and territorial documents for dentistry, medicine, nursing, pharmacy, occupational therapy, physiotherapy, psychology and veterinary medicine. Dentistry included two and nursing included nine specific pain competencies. No references to competencies related to pain were found in the remaining health science documents. In contrast, the national competency requirements for veterinary medicine, surveyed as a comparison, included nine pain competencies. All documents included competencies pertaining to ethics. The lack of competencies related to pain has implications for advancing skillful and ethical practice. The lack of attention to pain competencies limits the capacity of health care professionals to alleviate suffering, foster autonomy and use resources justly. Influencing professional bodies to increase the number of required entry-to-practice pain competencies may ultimately have the greatest impact on education and practice.


Prevalence of psychiatric disorders in a group of adult patients seeking general dental care.
To determine the prevalence of psychiatric disorders in adult patients seeking general dental care because their presence can have a significant impact on how these patients should be managed. Medical history questionnaires from 442 randomly selected patients who presented for examination at the Virginia Commonwealth University School of Dentistry were reviewed for a positive history of psychiatric conditions, whether they were being treated for their condition, and what medications were prescribed. Twenty percent of the patients had a positive history of a psychiatric disorder. The most common disorder for both sexes was depression. Other disorders included anxiety, bipolar disorder, eating disorder, claustrophobia, attention deficit disorder, seasonal affective disorder, and schizophrenia. More than one disorder was reported by 50% of men and 37% of women, the most common combination being depression and anxiety. Seventy-seven percent of women and 69% of men were under active treatment. Commonly prescribed medications, which can have important adverse effects, included selective serotonin uptake inhibitors, benzodiazepines, lithium, and tricyclic antidepressants. Twenty patients reported taking more than one medication for their disorder. A significant number of dental patients have a psychiatric disorder. Because such disorders can affect the patient's response to dental treatment and require treatment modifications, and the adverse effects of the medications being used can alter the oral environment, clinicians need to be aware of their presence and the proper way to manage these patients.


Returning research results to individuals who are incarcerated in the USA.
The return of research results to populations and individuals is increasingly recognised as both important but ethically complicated. In the USA, there are few studies or detailed evidence-based practices on the return of research results to individuals who are incarcerated. In general, return of research results is not required with some exceptions; however, there are reasons to believe that in many cases returning results is most consistent with the ethical conduct of research. With individuals who are incarcerated, specific considerations for this historically disadvantaged population should be addressed. These are privacy, therapeutic misconception, paternalism, actionability and communication. If research results are returned, the initial consent process should consider thoroughness regarding privacy rights, the researchers' role, clarity around next steps and communication methods. Prison leadership should be key stakeholders in the process to streamline communication. Returning research results can be a method to counter historical and current paternalism in the carceral setting, while shifting the culture on how research is understood and conducted in carceral settings. The decision to return results should balance benefits, risks and research ethics. Further research is needed to identify and implement optimal approaches for returning results in this population.


[Periodontal disease and general health--literature review].
Periodontal disease is a significant cause of alveolar bone resorption resulting ultimately in the loss of teeth. Inflammation of the periodontal tissues is initiated by bacteria of the oral micro-flora. Invading micro-organisms stimulate both protective and destructive inflammatory-immune responses involving cytokine release syndrome, chemokines, arachidonic acid metabolites, reactive oxygen and nitrogen intermediates, and matrix melloproteinases. The local infection may affect general health in two ways. First, transient bacteremia from the oral focus may result in metastatic infection in remote organs of susceptible hosts, such as bacterial endocarditis in patients with congenital or acquired heart diseases. Second, lipopolysaccharide and inflammatory mediators are not only involved in local tissue destruction but have the potential to modulate the course of cardiovascular, chronic obstructive lung and autoimmune diseases, diabetes mellitus and preterm birth. Epidemiologic observations, awaiting further verification by controlled prospective trials, underline the impact of oral health on general well-doing.


Cardiovascular and thermal responses to SCUBA diving.
Recreational SCUBA diving exposes individuals to environmental stresses not often encountered in other types of activity. These stresses include increased ambient pressure, raised partial pressure of O(2), increased resistance to movement, added weight and drag of diving equipment, cold stress, and a higher breathing resistance. One means to understand how such stresses affect a diver is to employ the stress-strain-adaptive response model. Physiologic adaptations, like an increase in VO(2) in response to cold stress, will minimize the strain placed on thermal balance. Nonphysiologic adaptive responses include those behavioral and equipment interventions that isolate the diver from a particular stress. Self-contained underwater breathing apparatus (SCUBA) isolates the diver from the inability to extract O(2) from the water; dive garments minimize the stress of cold water immersion. This review will focus on cardiorespiratory and thermal responses to SCUBA diving, using the stress-strain-adaptive response model to illustrate the interaction between diver and environment. Some responses like hyperventilation, cardiac arrhythmias, or cold injury due to vasoconstriction are not considered adaptive but are realistic possibilities in diving environments.


An institutional review of clear cell chondrosarcoma.
Clear cell chondrosarcoma is a rare bone neoplasm with a slow progressive clinical course and infrequent metastasis, but with a high local recurrence rate. We sought to ascertain the long-term outcome of patients with this neoplasm and to identify possible factors predicting survival. Sixteen patients with clear cell chondrosarcomas treated at one institution and who had long-term clinical followup were identified. All patients were treated by surgical resection of the tumor, which was classified as clear (> 2 mm) in 10 patients and marginal or intralesional in six patients. Three patients had local recurrence after a median of 1.7 years. Metastatic disease developed in four patients with a median time to diagnosis of 8.1 years. Ten-year overall survival of patients with clear cell chondrosarcomas was 89%, and disease-free survival was 68%. Patients with surgical resections comprising clear margins had longer disease-free survival compared with patients with marginal and intralesional tumors. Inadequate surgical resection of clear cell chondrosarcoma leads to risk of local recurrence and metastatic disease. The malignancy has a tendency to metastasize relatively late, therefore, followup of patients is necessary after the generally accepted 5-year period. Prognostic study, Level III (case control study). See the Guidelines for Authors for a complete description of levels of evidence.


Binding of beta 2-microglobulin by heterologous sera.
Purified human, rat or guinea-pig beta 2-microglobulin (beta 2m) was mixed with sera from guinea-pig, rat, mouse, rabbit, horse, goat, cow, rhesus monkey or man. The mixtures were incubated at 37 degrees C for various lengths of time. When the sera were separated by gel-chromatography on Sephadex G-200, beta 2m was traced not only in 'free' form but also in fractions with higher molecular weights. Evidence is presented suggesting that heterologous beta 2m binds to beta 2m-containing molecules in sera by exchange with the homologous counterpart.


A meta-analysis combining parallel and cross-over randomized controlled trials to assess impact of iodine fortified foods on urinary iodine concentration among children.
The purpose of this analysis was to combine evidence from parallel and cross-over randomized controlled trials to assess the impact of iodine fortified foods on urinary iodine concentration (UIC) in children. A structured search for studies on iodine intervention studies on MEDLINE, Pro Quest, and the Cochrane Library from Jan, 1990 to Dec, 2012 was carried out. Carry-over effect was estimated by general linear model. We explored two methods to pool continuous outcomes in a meta-analysis by combining parallel and cross-over trial designs. The standard mean difference was calculated for net change in UIC. Fixed or random-effects models were used to summaries fortified food response data. Meta-regression and covariate meta-analysis were performed to explore the influ-ence of confounders on the net pooled effect on UIC. The overall pooled estimate, which combined parallel with cross-over trials in the absence of carry-over effect of UIC from 9 studies, showed a significant increase in the fortified group compared with the control group (n=3448; standard mean difference=2.02 μg/L; 95% CI: 1.30, 2.73; I2=99%, τ2=1.81, p<0.01). Meta-regression analysis indicated that dose of the feeding was positively related to the effect size (regression coefficient=0.014; 95% CI: 0.003, 0.026; p<0.019). The net pooled effect size after removing the confounders was 1.59 (95% CI: 0.953, 2.23) μg/L. There was an association between intakes of io-dine fortified foods and UIC in children. These results suggest that we can combine parallel with cross-over trials for meta-analysis for nutrients such as iodine when absorption is high.


Electrochemical Potential Influences Phenazine Production, Electron Transfer and Consequently Electric Current Generation by Pseudomonas aeruginosa.
Pseudomonas aeruginosa has gained interest as a redox mediator (phenazines) producer in bioelectrochemical systems. Several biotic and abiotic factors influence the production of phenazines in synergy with the central virulence factors production regulation. It is, however, not clear how the electrochemical environment may influence the production and usage of phenazines by P. aeruginosa. We here determined the influence of the electrochemical potential on phenazine production and phenazine electron transfer capacity at selected applied potentials from -0.4 to +0.4 V (vs. Ag/AgClsat) using P. aeruginosa strain PA14. Our study reveals a profound influence of the electrochemical potential on the amount of phenazine-1-carboxylate production, whereby applied potentials that were more positive than the formal potential of this dominating phenazine (E° 'PCA = -0.24 V vs. Ag/AgClsat) stimulated more PCA production (94, 84, 128, and 140 μg mL-1 for -0.1, 0.1, 0.2, and 0.3 V, respectively) compared to more reduced potentials (38, 75, and 7 μg mL-1 for -0.4, -0.3, and -0.24 V, respectively). Interestingly, P. aeruginosa seems to produce an additional redox mediator (with E° ' ∼ 0.052 V) at applied potentials below 0 V, which is most likely adsorbed to the electrode or present on the cells forming the biofilm around electrodes. At fairly negative applied electrode potentials, both PCA and the unknown redox compound mediate cathodic current generation. This study provides important insights applicable in optimizing the BES conditions and cultures for effective production and utilization of P. aeruginosa phenazines. It further stimulates investigations into the physiological impacts of the electrochemical environment, which might be decisive in the application of phenazines for electron transfer with P. aeruginosa pure- or microbial mixed cultures.


Association of WISP1/CCN4 with Risk of Overweight and Gestational Diabetes Mellitus in Chinese Pregnant Women.
Obese women with gestational diabetes mellitus (GDM) have a higher risk of adverse outcomes than women with obesity or GDM alone. Our study is aimed at investigating the discriminatory power of circulatory Wnt1-inducible signaling pathway protein-1 (WISP1), a novel adipocytokine, on the copresence of prepregnancy overweight/obesity and GDM and at clarifying the relationship between the WISP1 level and clinical cardiometabolic parameters. A total of 313 participants were screened from a multicenter prospective prebirth cohort: Born in Shenyang Cohort Study (BISCS). Subjects were examined with a 2 × 2 factorial design for body mass index (BMI) ≥ 24 and GDM. Between 24 and 28 weeks of pregnancy, follow-up individuals underwent an OGTT and blood sampling for cardiometabolic characterization. We observed that the WISP1 levels were elevated in prepregnancy overweight/obesity patients with GDM, compared with nonoverweight subjects with normal blood glucose (3.45 ± 0.89 vs. 2.91 ± 0.75 ng/mL). Multilogistic regression analyses after adjustments for potential confounding factors revealed that WISP1 was a strong and independent risk factor for prepregnancy overweight/obesity with GDM (all ORs > 1). In addition, the results of the ROC analysis indicated that WISP1 exhibited the capability to identify individuals with prepregnancy overweight/obesity and GDM (all AUC > 0.5). Finally, univariate and multivariate linear regression showed that WISP1 level was positively and independently correlated with fasting blood glucose, systolic blood pressure, and aspartate aminotransferase and was negatively correlated with HDL-C and complement C1q. WISP1 may be critical for the prediction, diagnosis, and therapeutic strategies against obesity and GDM in pregnant women.


Hyaluronic acid increases tendon derived cell viability and collagen type I expression in vitro: Comparative study of four different Hyaluronic acid preparations by molecular weight.
Hyaluronic Acid (HA) has been already approved by Food and Drug Administration (FDA) for osteoarthritis (OA), while its use in the treatment of tendinopathy is still debated. The aim of this study was to evaluate in human rotator cuff tendon derived cells the effects of four different HA on cell viability, proliferation, apoptosis and the expression of collagen type I and collagen type III. An in vitro model was developed on human tendon derived cells from rotator cuff tears to study the effects of four different HA preparations (Ps) (sodium hyaluronate MW: 500-730 KDa - Hyalgan®, 1000 kDa Artrosulfur HA®, 1600 KDa Hyalubrix® and 2200 KDa Synolis-VA®) at various concentrations. Tendon derived cells morphology were evaluated after 0, 7 and 14 d of culture. Viability, proliferation, apoptosis were evaluated after 0, 24 and 48 h of culture. The expression and deposition of collagen type I and collagen type III were evaluated after 1, 7 and 14 d of culture. All HAPs tested increased viability and proliferation, in dose dependent manner. HAPs already reduce apoptosis at 24 h compared to control cells (without HAPs). Furthermore, HAPs stimulated the synthesis of collagen type I in a dose dependent fashion over 14 d, without increase in collagen type III; moreover, in the presence of Synolis-VA® the expression and deposition of collagen type I was significantly higher as compare with the other HAPs. HAPs enhanced viability, proliferation and expression of collagen type I in tendon derived cells.


The effect of Pediococcus acidilactici on the gut microbiota and immune status of on-growing red tilapia (Oreochromis niloticus).
To assess Pediococcus acidilactici as a dietary supplement for on-growing red tilapia (Oreochromis niloticus). Tilapia were fed either a control diet or control diet supplemented with Ped. acidilactici at 10(7) CFU g(-1) for 32 days. Ped. acidilactici colonized the intestinal tract and significantly affected the intestinal microbial communities. PCR-DGGE revealed direct antagonism of gastric Ped. acidilactici with an endogenous uncultured bacterium during a period of reverting to nonsupplemented feeding. Light microscopy revealed that gut integrity and leucocyte levels were unaffected by Ped. acidilactici; however, blood leucocyte levels and serum lysozyme activity were elevated after 14-days' feeding. No significant improvements in growth performance were observed at the end of the trial (day 32), but survival was significantly higher in the probiotic group. The study demonstrates that oral supplementation of Ped. acidilactici modulates intestinal bacterial communities in on-growing red tilapia and also stimulates some aspects of the nonspecific immune response. To our knowledge this is the first study assessing the effects of probiotics on the gut microbiota of tilapia using culture-independent methods. Such methods are crucial to understand the mechanisms which underpin and mediate host benefits.


Carbon monoxide release from ultrasound-sensitive microbubbles improves endothelial cell growth.
Carbon monoxide is a gasotransmitter that may be beneficial for vascular tissue engineering and regenerative medicine strategies because it can promote endothelial cell (EC) proliferation and migration by binding to heme-containing compounds within cells. For example, CO may be beneficial for vascular cognitive impairment and dementia because many patients' disrupted blood-brain barriers do not heal naturally. However, control of the CO dose is critical, and new controlled delivery methods need to be developed. This study developed ultrasound-sensitive microbubbles with a carefully controlled precipitation technique, loaded them with CO, and assessed their ability to promote EC proliferation and function. Microbubbles fabricated with perfluoropentane exhibited good stability at room temperature, but they could still be ruptured and release CO in culture with application of ultrasound. Microbubbles synthesized from the higher boiling point compound, perfluorohexane, were too stable at physiological temperature. The lower-boiling point perfluoropentane microbubbles had good biocompatibility and appeared to improve VE-cadherin expression when CO was loaded in the bubbles. Finally, tissue phantoms were used to show that an imaging ultrasound probe can efficiently rupture the microbubbles and that the CO-loaded microbubbles can improve EC spreading and proliferation compared to control conditions without microbubbles as well as microbubbles without application of ultrasound. Overall, this study demonstrated the potential for use of these ultrasound-sensitive microbubbles for improving blood vessel endothelialization.


Large Stroke Vertical PZT Microactuator With High-Speed Rotational Scanning.
A thin-film piezoelectric microactuator using a novel combination of active vertical translational scanning and passive resonant rotational scanning is presented. Thin-film lead-zirconate-titanate unimorph bending beams surrounding a central platform provide nearly 200-μm displacement at 18 V with bandwidth greater than 200 Hz. Inside the platform, a mirror mount, or mirror surface, supported by silicon dioxide spring beams can be excited to resonance by low-voltage; high-frequency excitation of the outer PZT beams. Over ±5.5° mechanical resonance is obtained at 3.8 kHz and ±2 V. The combination of large translational vertical displacements and high-speed rotational scanning is intended to support real-time cross-sectional imaging in a dual axes confocal endomicroscope.


Molecular evidence for multiple paternity in a feral population of green swordtails.
Genetic parentage analyses provide insights into mating systems and have revealed widespread evidence for polyandry in natural populations. Here, we use 5 microsatellite markers to estimate female mating rates in a feral population of green swordtails, Xiphophorus helleri, a live-bearing poeciliid fish that has become a model system in the study of precopulatory mate choice and mating competition. Although heralded as a potential model for investigating sperm competition as early as 1950, there has been no attempt to explore postcopulatory sexual selection in its mating system. We thus obtained information on the prevalence, and therefore biological relevance, of polyandry from a wild population. We genotyped the offspring from 14 wild-caught gravid females and determined the number of fathers in each brood using allele counting methods and the programs GERUD and PARENTAGE. Our analyses revealed that 57% (allele counts and GERUD) and 71% (PARENTAGE) of the sampled broods had at least 2 sires, with a global mean 1.74 fathers per brood. Paternity skew was generally high in mixed paternity broods so that our analyses almost certainly underestimate actual mating frequencies in the wild. Our data provide a solid underpinning for future studies of postcopulatory sexual selection in this species.


Physical injury risks associated with drinking water arsenic treatment.
We estimated the number of transportation deaths that would be associated with water treatment in Albuquerque to meet the EPA's recently proposed revisions of the Maximum Contaminant Level (MCL) for arsenic. Vehicle mileage was estimated for ion exchange, activated alumina, and iron coagulation/microfiltration water treatment processes to meet an MCL of 0.020 mg/L, 0.010 mg/L, 0.005 mg/L, and 0.003 mg/L. Local crash, injury, and death rates per million vehicle miles were used to estimate the number of injuries and deaths. Depending on the water treatment options chosen, we estimate that meeting an arsenic MCL of 0.005 mg/L will result in 143 to 237 crashes, 58 to 98 injuries, and 0.6 to 2.6 deaths in Albuquerque over a 70-year period, resulting in 26 to 113 years of life lost. The anticipated health benefits for Albuquerque residents from a 0.005 mg/L arsenic MCL, estimated using either a multistage Weibull or Poisson model, ranged from 3 to 80 arsenic-related bladder and lung cancer deaths prevented over a 70-year period, adding between 43 and 1,123 years of life. Whether a revised arsenic MCL increases or reduces overall loss of life in Albuquerque depends on the accuracy of EPA's cancer risk assessment. If the multistage Weibull model accurately estimates the benefits, the years of life added is comparable or lower than the anticipated years lost due to transportation associated with the delivery of chemicals, disposal of treatment waste, and operation of the water treatment system. Coagulation/microfiltration treatment will result in substantially fewer transportation deaths than either ion exchange or activated alumina.


Oral Gastrografin Facilitates Delineation of Intestinal Tracts in CT-Based Brachytherapy for Uterine Cervical Cancer.
Three-dimensional image-guided brachytherapy (3D-IGBT) using computed tomography (CT) is an essential component of definitive radiation therapy for uterine cervical cancer (UCC). Treatment planning for CT-based 3D-IGBT requires delineating the high-risk clinical target volume (CTVHR) and the organs at risk (OARs), which is difficult when the small intestine is adjacent to those delineation targets. Uncertainty in target delineation threatens the validity of 3D-IGBT treatment plans. To address this issue, we introduce the use of diatrizoate meglumine and diatrizoate sodium (gastrografin), an orally administrable iodine-based radiopaque contrast agent. We present two cases of UCC treated with CT-based 3D-IGBT and describe how intraluminal enhancement of the small intestine by oral gastrografin pretreatment facilitated discrimination between the small intestine and the adjacent CTVHR (case no.1) or the rectosigmoid colon (case no. 2). Oral gastrografin pretreatment is a simple and cost-effective method that allows distinguishing the small intestine from the adjacent delineation target (i.e., CTVHR and the OARs) in CT-based 3D-IGBT for UCC.


Regulation of activity and localization of the WNK1 protein kinase by hyperosmotic stress.
Mutations within the WNK1 (with-no-K[Lys] kinase-1) gene cause Gordon's hypertension syndrome. Little is known about how WNK1 is regulated. We demonstrate that WNK1 is rapidly activated and phosphorylated at multiple residues after exposure of cells to hyperosmotic conditions and that activation is mediated by the phosphorylation of its T-loop Ser382 residue, possibly triggered by a transautophosphorylation reaction. Activation of WNK1 coincides with the phosphorylation and activation of two WNK1 substrates, namely, the protein kinases STE20/SPS1-related proline alanine-rich kinase (SPAK) and oxidative stress response kinase-1 (OSR1). Small interfering RNA depletion of WNK1 impairs SPAK/OSR1 activity and phosphorylation of residues targeted by WNK1. Hyperosmotic stress induces rapid redistribution of WNK1 from the cytosol to vesicular structures that may comprise trans-Golgi network (TGN)/recycling endosomes, as they display rapid movement, colocalize with clathrin, adaptor protein complex 1 (AP-1), and TGN46, but not the AP-2 plasma membrane-coated pit marker nor the endosomal markers EEA1, Hrs, and LAMP1. Mutational analysis suggests that the WNK1 C-terminal noncatalytic domain mediates vesicle localization. Our observations shed light on the mechanism by which WNK1 is regulated by hyperosmotic stress.


Derivatization for electrospray ionization mass spectrometry. 3. Electrochemically ionizable derivatives.
In this paper, the use of ferrocene-based "electrochemically ionizable" derivatives to enhance ES-MS analysis of simple alcohols, sterols, and phenols is discussed. These derivatives are designed to take advantage of the electrolysis process inherent to operation of the ES ion source for selective ionization. Derivatization procedures, electrochemical character of the derivatives, and the ES-MS operational parameters necessary to maximize electrochemical ionization and to enhance gas-phase detection are presented with reference to ferrocenecarbamate ester derivatives of a variety of alcohol standards, as well as the ferroceneboronate derivative of the diol, pinacol. Tandem mass spectrometric analysis of the derivatives (precursor and product ion spectra) is shown to provide derivative confirmation, enhanced detection, and additional analyte structure information. The utility of this derivatization approach for the selective detection of alcohols in complicated mixtures is demonstrated using a saw palmetto (Serenoa repens) fruit extract known to contain a variety of alcohols at low levels.