Osteogenic stem cell selection for repair and regeneration.
The first osteogenic cells to attach to a titanium (Ti) implant after placement are the multipotent stromal cells (MSCs) that circulate in the bloodstream and are recruited to the site of tissue damage. The reservoirs of these cells are heterogeneous in nature, consisting of a mixture of cells with varying differentiation abilities. In order to utilise these cells and to reduce the chance of unwanted events during regenerative therapies, the selection of a subset of cells that is truly multipotent is required. The behaviour of these cells has been shown to be altered by modifications to Ti implant surfaces, most notably rough, hydrophilic Ti. These changes in behaviour underpin the differences seen in clinical performance of these surfaces. In this study Human bone marrow derived stromal cells (hBMSCs) have been cultured on modified Ti surfaces in order to analyse these changes in cell behaviour. The results demonstrate the different effects of the surfaces and suggest that one surface selectively enriches the population with osteogenic adult 'stem cells' by inducing the cell death of the more differentiated cells. Combined with subsequent expansion in bioreactors before implantation, this may lead to a new source of cells for regenerative therapies.


Simultaneous and interleaved acquisition of NMR signals from different nuclei with a clinical MRI scanner.
Modification of a clinical MRI scanner to enable simultaneous or rapid interleaved acquisition of signals from two different nuclei. A device was developed to modify the local oscillator signal fed to the receive channel(s) of an MRI console. This enables external modification of the frequency at which the receiver is sensitive and rapid switching between different frequencies. Use of the device was demonstrated with interleaved and simultaneous 31 P and 1 H spectroscopic acquisitions, and with interleaved 31 P and 1 H imaging. Signal amplitudes and signal-to-noise ratios were found to be unchanged for the modified system, compared with data acquired with the MRI system in the standard configuration. Interleaved and simultaneous 1 H and 31 P signal acquisition was successfully demonstrated with a clinical MRI scanner, with only minor modification of the RF architecture. While demonstrated with 31 P, the modification is applicable to any detectable nucleus without further modification, enabling a wide range of simultaneous and interleaved experiments to be performed within a clinical setting. Magn Reson Med 76:1636-1641, 2016. © 2015 The Authors. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.


Optimality Models and the Propensity Interpretation of Fitness.
The propensity account of fitness intends to solve the classical tautologicity issue by identifying fitness with a disposition, the ability to survive and reproduce. As proponents recognized early on, this account requires operational independence from actual reproductive success to avoid circularity and vacuousness charges. They suggested that operational independence is achieved by measuring fitness values through optimality models. Our goal in this article is to develop this suggestion. We show that one plausible procedure by which these independent operationalizations could be thought to take place, and which is in accordance with what is said in the optimality literature, is unsound. We provide two independent lines of reasoning to show this. We then provide a sketch of a more adequate account of the role of optimality models in evolutionary contexts and draw some consequences.


Assessment of right and left ventricular function by tissue Doppler echocardiography in patients after biventricular repair of pulmonary atresia with intact ventricular septum.
Survival of patients with PAIVS has improved significantly with adoption of a selective management approach based on RV and coronary arterial anatomy. We sought to determine the right (RV) and left ventricular (LV) function of patients with pulmonary atresia and intact ventricular septum (PAIVS) long-term after biventricular repair. The regional ventricular function of 22 patients (11 males) aged 12.8+/-5.6 years with PAIVS after biventricular repair was assessed by tissue Doppler echocardiography (TDE). The results were compared to those of 22 age-matched controls. When compared with controls, patients had significantly reduced myocardial tissue velocities, systolic and diastolic strain rate, and peak systolic strain in the basal, mid-, and apical segments of the RV free wall, interventricular septum and LV lateral wall (all with p<0.01). Heterogeneity of systolic and diastolic strain rate was observed in the RV free wall and interventricular septum in patients but not in controls. Impairment of RV and LV regional longitudinal myocardial function occurs in patients with PAIVS late after biventricular repair.


Early postoperative transtibial articular fistula formation after anterior cruciate ligament reconstruction: a review of three cases.
This is a retrospective case report of three cases with an early postoperative transtibial fistula after anterior cruciate ligament reconstruction (ACL). The patients had undergone ACL reconstruction and complained of fluid drainage through the not-healed wound or swelling localized on the anteromedial aspect of the ipsilateral proximal tibia during the early postoperative. Magnetic resonance imaging showed a multilocular fluid-filled cyst arising from the distal hole of the tibial bone tunnel. Open resection of the fistula and the cyst was performed in all cases and communication between the tibial tunnel and the joint space was confirmed. During revision surgery the distal hole of the tibial tunnel was covered with a fascio-periosteal flap. All wounds healed without complications. There was no recurrence of drainage or cyst formation. At 2 years follow-up the knee function was normal and was not affected by the complication in any of the patients. Early postoperative transtibial fistulae after ACL reconstruction are rare complications that clinically present either as anterior tibial cysts or persistent wound drainage. Surgical treatment is required, and some delay in the rehabilitation routine is required, but the final outcome is not affected.


Factors confounding cytosolic calcium measurements in Jurkat E6.1 cells during exposure to ELF magnetic fields.
Reported changes in the cytosolic calcium concentration ([Ca2+](c)) as a result of exposure to extremely low frequency (ELF) magnetic fields (MF) have been equivocal. In this study, we examine the possibility that some of these differences are attributable to variability associated with the cell cycle, pH of the suspension medium, and response to a calcium agonist. We used a custom designed spectrofluorimeter to measure [Ca2+](c) in Indo 1-AM loaded Jurkat E6.1 cells suspended in conditioned RPMI 1640 medium containing 10% fetal bovine serum. Four exposures were examined: zero static MF (Null), 60 Hz 100 microT(peak) sinusoidal MF (AC), 78 microT static MF (DC), and the combination of the 60 Hz and the 78 microT static MF (AD + DC). A significant decrease in normalized [Ca2+](c) values between 375-495 s for the DC and AC + DC groups was found in comparison to the Null group. However, statistical analysis indicated that cell cycle and quality of the alpha-CD3 monoclonal antibody response were significant covariates, while pH was not a significant covariate. When the effect of these covariates was taken into account, all exposure groups were significantly different from the control. Our results suggest that ELF MF effects may not be seen unless correction is made for biological variability of each cell preparation with respect to cell cycle and [Ca2+](c) response to antigen stimulation.


The effects of repeated dermal application of capsaicin to the human skin on pain and vasodilatation induced by intradermal injection of acid and hypertonic solutions.
1. The effect of repeated capsaicin application on the skin of the volar surface of the forearm on the pain sensation and on the increase in blood flow induced by intradermal injection of low pH media or hypertonic solutions was investigated in 13 healthy volunteers. 2. Low pH media (4, and 2.5) were obtained by adding HCl to 0.9% saline. Hypertonic solutions (300 and 600 mM) were obtained by adding NaCl to pH 7.4, 0.9% saline. Capsaicin (1% in 50% ethanol) was painted on the volar skin of one forearm, chosen at random, for 7 days. The contralateral forearm was treated with the capsaicin vehicle. Pain was assessed by a visual analogue scale and skin blood flow by a laser doppler flowmeter. 3. Pain sensation and increase in blood flow (both peak and area under the curve) induced by low pH media were markedly reduced in the capsaicin pretreated side. Capsaicin pretreatment also reduced the increase in blood flow, but did not affect the pain response induced by hypertonic saline solutions. 4. Repeated application of capsaicin to the human skin inhibits both the sensory (pain) and 'efferent' (vasodilatation) responses induced by low pH media, whereas it reduces the vasodilatation, but not the pain caused by hypertonic media. 5. Repeated application of capsaicin to the human skin, a therapy used in various diseases, discriminates between sensory, but not 'efferent' responses induced by different stimuli.


Hormones in the translational control of early germination in wheat embryos.
Gibberellic and abscisic acid seem to exert their effects on germination by acting in some way at the level of translation. The two hormones may act at different sites.


Ocular findings in polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy.
A routine ophthalmologic examination supplemented by color fundus photography and, in 2 cases, by fluorescein angiography was performed in 8 patients with PLO-SL (= polycystic lipomembranous dysplasia with sclerosing leukoencephalopathy). Five patients showed some degree of retinal nerve fiber defect. The relation of the changes in ocular fundi to the pathological findings of PLO-SL are discussed.


Congenital absence of the pericardium: displacement of the heart associated with tricuspid insufficiency.
This report describes a patient with congenital absence of the left pericardium with displacement of the heart into the left pleural space and associated tricuspid insufficiency. Excision of the ruptured chordae, plication of the flail anterior leaflet, and tricuspid annuloplasty rendered a competent tricuspid valve.


Nontumorous vascular malformations in the liver: color Doppler ultrasonographic findings.
To investigate color Doppler and spectral wave characteristics of nontumorous vascular malformations in the liver. From September 1995 to January 2001, 32 cases of vascular malformations were identified by means of color Doppler ultrasonography and spectral wave analysis. Computed tomography, angiography, or both were performed in all cases. Five arterioportal and 14 portovenous malformations, 1 arteriovenous malformation, and 4 portoportal and 8 venovenous shunts were detected. Associations with Rendu-Osler-Weber syndrome in 6 cases and with cirrhotic liver in 12 cases were found Fourteen patients were liver disease free. In 3 cases, interventional procedures were necessary to reduce portal hypertension or cardiac dysfunction. The incidence of finding vascular malformations in 12,000 patients was 0.1%. Nontumorous vascular malformations are rare disorders in the liver. They may appear in patients with healthy livers and in patients with portal hypertension. Color Doppler ultrasonography and spectral wave analysis are capable of showing and differentiating different types of hepatic vascular malformations.


The chronic syndromes after previous treatment of pituitary tumours.
Ultimately, almost all patients who are appropriately treated for pituitary tumours enter a chronic phase with control or cure of hormonal excess, adequate treatment of pituitary insufficiency and relief of mass effects. This phase is associated with improvement of initial signs and symptoms, but also with the persistent consequences of the initial disease and associated treatments. Pituitary insufficiency is a common denominator in many of these patients, and is associated with a reduction in quality of life, despite adequate endocrine substitution. Hypothalamic dysfunction can be present in patients previously treated for visual impairments caused by large suprasellar adenomas, or craniopharyngiomas. In addition to hypopituitarism, these patients can have multisystem morbidities caused by altered hypothalamic function, including weight gain and disturbed regulation of sleep-wake cycles. Mortality can also be affected. Patients cured of Cushing disease or acromegaly have chronic multisystem morbidities (in the case of Cushing disease, also affecting mortality) caused by irreversible effects of the previous excesses of cortisol in Cushing disease and growth hormone and insulin-like growth factor 1 in acromegaly. In addition to early diagnosis and treatment of pituitary tumours, research should focus on the amenability of these chronic post-treatment syndromes to therapeutic intervention, to improve quality of life and clinical outcomes.


Enzyme-linked immunosorbent assay detection of prostate-specific antigen messenger ribonucleic acid in prostate cancer.
To develop a rapid, sensitive, reverse transcriptase-polymerase chain reaction (RT-PCR) prostate-specific antigen (PSA) messenger ribonucleic acid (mRNA) detection method by applying colorimetric enzyme-linked immunosorbent assay (ELISA). Total RNA was extracted from 16 urogenital cancer cells (including PSA-producing LNCaP cells) from pelvic and inguinal lymph node aspiration biopsy samples from patients with prostate, bladder, and penile cancer, as well as from blood samples of 500 patients with urogenital cancer. We used rTth polymerase for RT and PCR. The RNA target was amplified by RT-PCR with dinitrophenyl-labeled primer. The PCR product was denatured and hybridized on a PSA-specific probe-coated microwell plate. In 1 6 cancer cell lines, only LNCaP cells expressed especially high PSA mRNA values, with an optical density (OD) greater than 3. In other cell lines, two testicular cells had relatively high ODs, 1.909 and 0.987, respectively. A high PSA mRNA value was obtained by fine needle aspiration from pelvic lymph node specimens of cytologically positive lymph nodes from patients with prostate cancer but not from patients with cytologically proved bladder or penile cancer. Sensitivity and specificity of fine needle aspiration samples were 70% and 100%, respectively. Blood tests obtained from patients with prostate cancer demonstrated high PSA mRNA values. The PSA mRNA RT-PCR ELISA method provides a sensitive photometric enzyme immunoassay for the detection of PSA mRNA, using nonradioactive techniques.


Effects of spatial heterogeneity on bacterial genetic circuits.
Intracellular spatial heterogeneity is frequently observed in bacteria, where the chromosome occupies part of the cell's volume and a circuit's DNA often localizes within the cell. How this heterogeneity affects core processes and genetic circuits is still poorly understood. In fact, commonly used ordinary differential equation (ODE) models of genetic circuits assume a well-mixed ensemble of molecules and, as such, do not capture spatial aspects. Reaction-diffusion partial differential equation (PDE) models have been only occasionally used since they are difficult to integrate and do not provide mechanistic understanding of the effects of spatial heterogeneity. In this paper, we derive a reduced ODE model that captures spatial effects, yet has the same dimension as commonly used well-mixed models. In particular, the only difference with respect to a well-mixed ODE model is that the association rate constant of binding reactions is multiplied by a coefficient, which we refer to as the binding correction factor (BCF). The BCF depends on the size of interacting molecules and on their location when fixed in space and it is equal to unity in a well-mixed ODE model. The BCF can be used to investigate how spatial heterogeneity affects the behavior of core processes and genetic circuits. Specifically, our reduced model indicates that transcription and its regulation are more effective for genes located at the cell poles than for genes located on the chromosome. The extent of these effects depends on the value of the BCF, which we found to be close to unity. For translation, the value of the BCF is always greater than unity, it increases with mRNA size, and, with biologically relevant parameters, is substantially larger than unity. Our model has broad validity, has the same dimension as a well-mixed model, yet it incorporates spatial heterogeneity. This simple-to-use model can be used to both analyze and design genetic circuits while accounting for spatial intracellular effects.


Anatomical brain connectivity can assess cognitive dysfunction in multiple sclerosis.
Brain disconnection plays a major role in determining cognitive disabilities in multiple sclerosis (MS). We recently developed a novel diffusion-weighted magnetic resonance imaging (DW-MRI) tractography approach, namely anatomical connectivitity mapping (ACM), that quantifies structural brain connectivity. Use of ACM to assess structural connectivity modifications in MS brains and ascertain their relationship with the patients' Paced-Auditory-Serial-Addition-Test (PASAT) scores. Relapsing-remitting MS (RRMS) patients (n = 25) and controls (n = 25) underwent MRI at 3T, including conventional images, T1-weighted volumes and DW-MRI. Volumetric scans were coregistered to fractional anisotropy (FA) images, to obtain parenchymal FA maps for both white and grey matter. We initiated probabilistic tractography from all parenchymal voxels, obtaining ACM maps by counting the number of streamlines passing through each voxel, then normalizing by the total number of streamlines initiated. The ACM maps were transformed into standard space, for statistical use. RRMS patients had reduced grey matter volume and FA, consistent with previous literature. Also, we showed reduced ACM in the thalamus and in the head of the caudate nucleus, bilaterally. In our RRMS patients, ACM was associated with PASAT scores in the corpus callosum, right hippocampus and cerebellum. ACM opens a new perspective, clarifying the contribution of anatomical brain disconnection to clinical disabilities in MS.


Cognitive disorders of infantile autism: a study employing the WISC, spatial relationship conceptualization, and gesture imitations.
Sixteen autistic children with WISC Performance IQs of 70 or above were analyzed to determine their conceptions of spatial relations, size comparisons, and gesture imitations through the use of the WISC, an originally devised Language Decoding Test (LDT), and a modified Gesture Imitation Test (GIT). WISC results were replicated as in previous studies. The autistic children showed an inability to acquire concepts of size comparison and spatial relationships through verbal instructions. They often gave peculiar responses (partial imitations), which seem to be related to their inability to integrate another person's body as a whole through visual input. These findings support the notion that the basic cognitive deficit is an impairment of symbolic-representational functioning, including language and body images, which results from a combination of delay and deviation of the symbolic-representational function.


Optimization and validation of RP-HPLC method for simultaneous estimation of palbociclib and letrozole.
A simple, rapid, and robust RP-HPLC method have been developed and validated to measure palbociclib (PB) and letrozole (LT) at single wavelength (254 nm). A isocratic elution of samples performed on Intersil C8 (4.6 mm × 250 mm particle size 5 μm) column with mobile phase consisting 0.02 M sodium dihydrogen phosphate buffer (pH 5.5): acetonitrile: methanol (80:10:10 v/v/v) delivered at flow rate 1.0 mL min-1. A good linear response was achieved over the range of 5-50 μg mL-1. The LODs for PB and LT were found to be 0.098 and 0.0821 µg mL-1, while the LOQs for PB and LT were 0.381-0.315 µg mL-1, respectively. The method was quantitatively evaluated in terms of system suitability test, linearity, precision, accuracy (recovery) and robustness as per standard guidelines. The method is simple, convenient and suitable for the analysis of PB and LT in bulk drug.


Immunohistochemical localization of creatine kinase BB in primary breast cancer: correlation with estrogen receptor content.
The brain-type (BB) isoenzyme of creatine kinase (CK) has recently been shown to be estrogen-regulated in human breast cancer cells in vitro. In this study we have correlated the presence of CK-BB, evaluated by immunoperoxidase procedures, with the estrogen receptor (ER) content in 35 primary breast cancers. Of 35 tumors examined, 66% revealed moderate to strong CK-BB immunoreactivity. Staining was exclusively located in the cytoplasm of neoplastic cells. A positive relationship was observed between CK-BB positivity and ER content with ER-rich tumors (greater than 50 fmoles/mg protein) showing a more intense immunoreactivity than ER-poor ones. The results indicate that CK-BB is estrogen-regulated in human breast cancer and suggest that evaluation of this enzyme may be of potential value for the detection of hormone responsive tumors.


Exploring skeletal muscle tolerance and whole-body metabolic effects of FDA-approved drugs in a volumetric muscle loss model.
Volumetric muscle loss (VML) is associated with persistent functional impairment due to a lack of de novo muscle regeneration. As mechanisms driving the lack of regeneration continue to be established, adjunctive pharmaceuticals to address the pathophysiology of the remaining muscle may offer partial remediation. Studies were designed to evaluate the tolerance and efficacy of two FDA-approved pharmaceutical modalities to address the pathophysiology of the remaining muscle tissue after VML injury: (1) nintedanib (an anti-fibrotic) and (2) combined formoterol and leucine (myogenic promoters). Tolerance was first established by testing low- and high-dosage effects on uninjured skeletal muscle mass and myofiber cross-sectional area in adult male C57BL/6J mice. Next, tolerated doses of the two pharmaceutical modalities were tested in VML-injured adult male C57BL/6J mice after an 8-week treatment period for their ability to modulate muscle strength and whole-body metabolism. The most salient findings indicate that formoterol plus leucine mitigated the loss in muscle mass, myofiber number, whole-body lipid oxidation, and muscle strength, and resulted in a higher whole-body metabolic rate (p ≤ 0.016); nintedanib did not exacerbate or correct aspects of the muscle pathophysiology after VML. This supports ongoing optimization efforts, including scale-up evaluations of formoterol treatment in large animal models of VML.


The Deep Brain Stimulation Impairment Scale: A useful complement in assessment of well-being and functioning in DBS-patients - Results from a large multicentre survey in patients with Parkinson's disease.
Deep Brain Stimulation (DBS) has been proven to alleviate motor symptoms in Parkinson's Disease (PD). Regarding non-motor symptoms, however, inconsistencies have been reported, on whether DBS causes reductions in well-being and functioning. To assess motor and non-motor impairment in DBS-patients, the Deep Brain Stimulation Impairment Scale (DBS-IS) has been developed. Yet, the extent to which the DBS-IS detects impairment in DBS-patients and thus could serve as a useful tool that complements the PDQ-39 (gold standard) in assessment of well-being and functioning in PD-patients has not been shown. By comparing DBS and non-DBS-patients we aimed to identify DBS-specific symptoms. We thereby aimed to show in how far the DBS-IS complements the PDQ-39 in assessing well-being and functioning in PD patients under DBS. In a cross-sectional study, 186 DBS-patients were matched (for age, disease duration and sex) to 186 non-DBS-patients (N = 372) and the two groups were compared regarding well-being and functioning: Impairment was assessed via DBS-IS and overall Quality of Life (QoL) was assessed via PDQ-39. Additionally, we analyzed differences in impairment between age and disease duration clusters. DBS-patients showed significantly higher total impairment (DBS-IS) and significantly higher impairment on the subscales Postural Instability and Gait difficulties and speaking difficulties than non-DBS-patients. Impairment increased with age and disease duration and, overall, differences in impairment rose by age. Overall QoL (PDQ-39) was non-significantly lower in DBS-patients. Since there is evidence that the PDQ-39 misses some DBS-specific symptoms, the DBS-IS is recommended to complement the PDQ-39 when assessing DBS-patients.


Surgical treatment result of abdominal aortic aneurysm in Behçet's disease.
We report our surgical treatment results of abdominal aortic aneurysm (AAA) in Behçet's disease patient. Between September 1998 and June 2006, the authors have performed 21 procedures for AAA in 12 patients with Behçet's disease. Male to female sex ratio was 3:1 and mean age was 34 years old. Behçet's disease was diagnosed clinically using criteria of International Study Group for Behçet's Disease (1990). Retrospective analysis was made. There were six infrarenal, five suprarenal, and one double (suprarenal and infrarenal)AAA. Six graft interposition, six patch closure, and one stent-graft insertion were performed (one graft interposition and one patch closure were simultaneously performed for double AAA). Eight recurrent aneurysms were noted in six (50%) patients. Four stent-graft insertion, two patch closures, one graft interposition and one explothoracotomy only were performed for recurrent aneurysms. Overall recurrence rate of 21 procedures was 38.1%; 14.3% for graft interposition, 62.5% for patch closure, and 40% for stent-graft insertion. Though the resection and graft interposition is technically difficult in many occasions, it should be considered as the procedure of choice for abdominal aortic aneurysm in Behçet's disease. Endovascular interventions may be one of the treatment modality but the result needs further long-term follow-up.


An Fc variant with two mutations confers prolonged serum half-life and enhanced effector functions on IgG antibodies.
The pH-selective interaction between the immunoglobulin G (IgG) fragment crystallizable region (Fc region) and the neonatal Fc receptor (FcRn) is critical for prolonging the circulating half-lives of IgG molecules through intracellular trafficking and recycling. By using directed evolution, we successfully identified Fc mutations that improve the pH-dependent binding of human FcRn and prolong the serum persistence of a model IgG antibody and an Fc-fusion protein. Strikingly, trastuzumab-PFc29 and aflibercept-PFc29, a model therapeutic IgG antibody and an Fc-fusion protein, respectively, when combined with our engineered Fc (Q311R/M428L), both exhibited significantly higher serum half-lives in human FcRn transgenic mice than their counterparts with wild-type Fc. Moreover, in a cynomolgus monkey model, trastuzumab-PFc29 displayed a superior pharmacokinetic profile to that of both trastuzumab-YTE and trastuzumab-LS, which contain the well-validated serum half-life extension Fcs YTE (M252Y/S254T/T256E) and LS (M428L/N434S), respectively. Furthermore, the introduction of two identified mutations of PFc29 (Q311R/M428L) into the model antibodies enhanced both complement-dependent cytotoxicity and antibody-dependent cell-mediated cytotoxicity activity, which are triggered by the association between IgG Fc and Fc binding ligands and are critical for clearing cancer cells. In addition, the effector functions could be turned off by combining the two mutations of PFc29 with effector function-silencing mutations, but the antibodies maintained their excellent pH-dependent human FcRn binding profile. We expect our Fc variants to be an excellent tool for enhancing the pharmacokinetic profiles and potencies of various therapeutic antibodies and Fc-fusion proteins.


Traditional bone setting service users and associated factors among people with trauma in Mecha district, Ethiopia.
World Health Organization stated that traditional medicine is an important part of health care and countries need to consider integrating it into their primary health care system. Traditional bone setting has a long history in Ethiopia and it enjoys enormous acceptance in the community. However, these methods are raw, there is no standardized training and at the same time, complications are common. Therefore, this research aimed to assess the prevalence of traditional bone setting service utilization and associated factors among people with trauma in Mecha district. Methods A Community- based cross-sectional study design was employed from January 15 to February 15, 2021. A total of 836 participants were selected using a simple random sampling technique. Binary and multiple logistic regressions were employed to assess the association between the independent variables with traditional bone setting service utilization. Results The prevalence of traditional bone setting service utilization was 46.05%. Factors significantly associated with TBS utilization were: Age ≥ 60 years (AOR = 0.13, 95% CI: 0.03- 0.43), rural residence (AOR = 3.63, 95% CI: 1.76 -7.50), occupation (merchant (AOR = 0.21, 95% CI: 0.07 -0.61), and housewife (AOR = 4.12, 95% CI: 1.33 -12.70), type of trauma: dislocation (AOR = 6.40, 95% CI: 3.69-11.10), and strain (AOR = 2.09, 95% CI: 1.05- 4.14)), site of trauma: extremity (AOR = 0.20, 95% CI: 0.11, 0.37), trunk (AOR = 0.08, 95% CI: 0.03-0.22), and shoulder (AOR = 0.20, 95% CI: 0.11-0.37), cause of trauma: fall down and natural deformity (AOR = 9.87, 95% CI: 5.93-16.42) and household annual income greater than > 36,500 (AOR = 2.33, 95% CI: 1.29-4.22). Conclusion The prevalence of traditional bone setting practice is high in the study area, despite recent advancements in the practice of orthopedics and trauma in Ethiopia. Since TBS services are more accepted in society, the integration of TBS into the health care delivery system is recommended.


Complying with the 2008 National Patient Safety Goals.
In 2003, the joint commission began publishing National Patient Safety Goals (NPSGs) and requiring accredited health care organizations to comply with these goals in an effort to reduce the number of medical errors. THE NPSGS are updated yearly with new requirements to promote specific improvements in patient safety. This article provides a review of the 2008 NPSGs and suggests ways in which information technology systems can address health care organizations' compliance with some of these goals.


Hydrolysis-resistant GTP analogs stimulate catecholamine release from digitonin-permeabilized PC12 cells.
The effect of the hydrolysis-resistant GTP analogs, guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) and guanylyl imidodiphosphate (GMPPNP), on norepinephrine (NE) secretion from digitonin-permeabilized rat pheochromocytoma cells, PC12, was examined. Although secretion in the presence of saturating Ca2+ (10 microM) was not affected by GTP gamma S or GMPPNP, secretion in the absence of Ca2+ was stimulated by these GTP analogs. Secretion induced by saturating concentrations of GTP gamma S or GMPPNP was approximately 80% of that induced by 10 microM Ca2+. Half-maximum stimulation was induced by 30 microM GTP gamma S or GMPPNP. Both Ca2(+)-stimulated and GTP gamma S-stimulated secretion were ATP dependent and inhibited by N-ethylmaleimide. The GTP gamma S-stimulated secretion of NE from permeabilized PC12 cells does not appear to result from either the release of Ca2+ or the activation of protein kinase C. Activation of protein kinase C by pretreatment of intact cells with 12-O-tetradecanoylphorbol 13-acetate caused a 50% increase in both Ca2(+)-stimulated and GTP gamma S-stimulated secretion. Cholera and pertussis toxins did not affect Ca2(+)-stimulated or GTP gamma S-stimulated NE secretion. Guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) and GTP inhibited GTP gamma S-stimulated secretion but not Ca2(+)-stimulated secretion. The inability of GDP beta S to inhibit Ca2(+)-stimulated secretion indicates that the process affected by GTP gamma S is not an essential step in the Ca2(+)-stimulated pathway.


Solid-pseudopapillary tumor of the pancreas: a surgical enigma?
Solid-pseudopapillary tumors (SPTs) of the pancreas have been reported as rare lesions with "low malignant potential" occurring mainly in young women. This study was designed to define the clinicopathological characteristics and the effect of surgical intervention. A retrospective review from January 1985 to July 2000 was performed. Clinicopathological, operative, and survival data were obtained. The Kaplan-Meier method and chi2 analysis were performed. All cases were re-reviewed by a senior pathologist. During this time, 24 patients were diagnosed as having SPTs (0.9%). Twenty females and four males were identified, with a median age of 39 years (range, 12-79). The median size of the lesions was 8.0 cm (range, 1-20). Two patients' tumors were found to be unresectable at initial presentation because of vascular invasion; both patients have remained alive with disease, one for 13 years and the other 1 year. At a median follow-up of 8 years, one recurrence occurred in 17 patients who underwent complete resection. Microscopic margin positive (P = .26), invasion of surrounding structures (P = .51), and size >5 cm (P = .20) were not significant predictors of survival. Four patients presented with synchronous liver metastasis and underwent resection of the primary tumor and the liver metastasis, with one patient dying of progression of metastatic disease at 8 months, another alive with recurrence in the liver at 6 years, and the last two alive without evidence of disease at 1 month and 11 years. SPT occurs predominantly in women (82%), although it can occur in men; all age groups are affected. Complete resection is associated with long-term survival even in the presence of metastatic disease.


Identification of oral bacteria on titanium implant surfaces by 16S rDNA sequencing.
To characterize the profile of microbial communities colonizing titanium implants with different surface treatments after exposure to the oral environment at the genus or higher taxonomic level. Sixteen titanium disks, machined or sandblasted large-grit and acid-etched (SLA), were mounted on removable intraoral splints worn by four patients. After 24 h of intraoral exposure, biofilm samples were collected from disks and supra/subgingival teeth areas. The 16S rDNA genes from each sample were amplified, sequenced with the Miseq Illumina instrument and analyzed. A total of 29 genera and seven more inclusive taxa, representing the phyla Firmicutes, Proteobacteria, Fusobacteria, Bacteroidetes, Actinobacteria and candidate division TM7 were identified in both titanium surfaces and teeth. No differences were found in relation to the operational taxonomic units (OTUs) and microbial diversity, assessed by Chao 1 and Shannon indices, when comparing SLA and machined titanium surfaces. Machined and SLA surfaces are colonized by similar numbers of prokaryotic OTUs after 24 h of exposure to the oral environment. Higher complexity of the titanium surface topography in the initial phase of biofilm maturation does not seem to significantly influence the colonizing microbiota.


Persistent non-homeostatic remodeling of aortic collagen following a brief episode of hypertension: A computational study.
The healthy adult aorta exhibits a remarkable homeostatic ability to respond to sustained changes in hemodynamic loads under many circumstances, but this mechanical homeostasis can be compromised or lost in natural aging and diverse pathological processes. Herein, we investigate persistent non-homeostatic changes in the composition and mechanical properties of the thoracic aorta in adult wild-type mice following 14 days of angiotensin II-induced hypertension. We employ a multiscale computational model of arterial growth and remodeling driven by mechanosensitive and angiotensin II-related cell signaling pathways. We find that experimentally observed findings can only be recapitulated computationally if the collagen deposited during the transient period of hypertension has altered properties (deposition stretch, fiber angle, crosslinking) compared with the collagen produced in the original homeostatic state. Some of these changes are predicted to persist for at least six months after blood pressure is restored to normal levels, consistent with the experimental findings.


Genetic and population structure of four Sardinian villages.
Data on microgeographic population structure on four neighbouring villages of Sardinia island (Italy) are presented and discussed. Two villages are located in the lowlands where malaria from Plasmodium falciparum was endemic until the eradication of paludism. The other two villages are located in the highlands and they were malaria-free because of the altitude. Census data, inbreeding, migration matrices and surname distributions have been collected. The genetic differentiation of the four villages, tested for 31 genetic polymorphisms (106 alleles), is only in part compatible with migration rates inferred from demographic data. The possible adaptive nature of some genetic markers with respect to malarial resistance is discussed. Ambiguous results from population genetics quantitative methods do not support definite answers.


Methicillin-resistant Staphylococcus aureus infection epidemiology and clinical response from tigecycline soft tissue infection trials.
Given increasing resistance, therapeutic options to treat MRSA soft tissue infections should be evaluated. This pooled analysis evaluated data from subjects enrolled in 6 tigecycline clinical trials with documented MRSA complicated skin and skin structure infections or diabetic foot infections (DFIs). Baseline characteristics were compared between subjects with and without molecularly classified community-acquired (CA) MRSA, specifically staphylococcal cassette chromosome mec (SCCmec) IV. Clinical response was compared by CA-MRSA designation and treatment group. A total of 378 subjects with MRSA soft tissue infections were identified, including 79 with DFI. A total of 249 (65.9%) were molecularly classified as CA-MRSA. Clinical response rates for MRSA soft tissue infection were similar between tigecycline and vancomycin (treatment difference, 1.0%; 95% confidence interval: -9.3, 12.0) as well as by infection type, SCCmec, and Panton-Valentine leukocidin (PVL) status. Tigecycline demonstrated comparable efficacy for treatment of MRSA soft tissue infections regardless of infection type, SCCmec, or PVL status.


Duplication and functional specialization of the telomere-capping protein Cdc13 in Candida species.
The budding yeast G-tail binding complex CST (Cdc13-Stn1-Ten1) is crucial for both telomere protection and replication. Previous studies revealed a family of Cdc13 orthologues (Cdc13A) in Candida species that are unusually small but are nevertheless responsible for G-tail binding and the regulation of telomere lengths and structures. Here we report the identification and characterization of a second family of Cdc13-like proteins in the Candida clade, named Cdc13B. Phylogenetic analysis and sequence alignment indicate that Cdc13B probably arose through gene duplication prior to Candida speciation. Like Cdc13A, Cdc13B appears to be essential. Deleting one copy each of the CDC13A and CDC13B genes caused a synergistic effect on aberrant telomere elongation and t-circle accumulation, suggesting that the two paralogues mediate overlapping and nonredundant functions in telomere regulation. Interestingly, Cdc13B utilizes its C-terminal OB-fold domain (OB4) to mediate self-association and binding to Cdc13A. Moreover, the stability of the heterodimer is evidently greater than that of either homodimer. Both the Cdc13 A/A homodimer and A/B heterodimer, but not the B/B homodimer, recognized the telomere G-tail repeat with high affinity and sequence specificity. Our results reveal novel evolutionary elaborations of the G-tail-binding protein in Saccharomycotina yeast, suggesting a drastic remodeling of CDC13 that entails gene duplication, fusion, and functional specialization. The repeated and independent duplication of G-tail-binding proteins such as Cdc13 and Pot1 hints at the evolutionary advantage of having multiple G-tail-binding proteins.


Histopathologic findings and diagnostic difficulties posed with use of escharotic agents for treatment of skin lesions: a case report and review of the literature.
Escharotic agents have been used as alternative therapy for treatment of skin cancer and skin problems for centuries. Internet web sites such as online health product stores and eBay have made them widely available to the general public. The use of these agents carries risk of incomplete removal of tumor, damage of surrounding healthy tissues and marked scarring with poor cosmetic outcome. We report the case of a 27-year-old man who presented with history of moles and self-treatment with an escharotic agent containing bloodroot in order to document the histopathologic findings of topical bloodroot treatment and to show the clinical consequences that may occur in the unsuspecting public. To the best of our knowledge, the histological features following use of bloodroot (Sanguinaria canadensis) have not yet been documented.


Prophylactic antibiotics in amputation of the lower extremity for ischemia. A placebo-controlled, randomized trial of cefoxitin.
Of 152 patients who were scheduled for an amputation for ischemia, seventy-seven were randomly assigned to perioperative prophylaxis with cefoxitin (Mefoxin) and seventy-five patients, to injections of a placebo. The patients were followed for twenty-one days or, in the case of wound complications, to the end of treatment. An infected wound occurred in 38.7 per cent of the patients in the placebo group and 16.9 per cent of those in the antibiotic group (p less than 0.005). Clostridial infection occurred in eight patients in the placebo group and in none in the antibiotic group (p = 0.003). Three of the patients with clostridial infection died of gas gangrene. A multivariate analysis showed that the absence of antibiotic prophylaxis increased the risk of infection by a factor of 3.3 (p = 0.004) and increased the need for reamputation by a factor of 4.5 (p = 0.003). We concluded that amputation patients should have prophylaxis with a broad-spectrum antibiotic given perioperatively.


Predictors of perceived health care quality for registered nurses during and after health care reform.
Limited research has focused on the predictive nature of organizational culture and trust on registered nurses' perceived health care quality in reformed health care systems. The purpose of this article was to investigate nurses' perceptions of organizational culture factors, trust in employer, and perceived health care quality during and 5 years after major organizational reform in the acute care setting and to test a model linking culture to perceived health care quality. Survey data collected from two samples of nurses (N = 222,343) during and 5 years after major organizational reform in the acute care setting of one Canadian province were analyzed, and an exploratory model linking aspects of culture, trust, and quality was tested. For both periods, most variable scores were in the low range and depicted moderately positive intercorrelations with each other. Support for the proposed model was mixed. Select culture variables predicted health care quality at both periods, but trust emerged as a significant predictor in 2000 only. The findings support the negative impact of system transformation on nurses and the link between culture and health care quality. The study findings suggest that managers and policy makers must develop and implement supportive and nurturing strategies that will enhance the organizational culture (emotional climate, collaborative relations), which should result in more positive perceptions of health care quality. However, further research is required to gain a better understanding of the relationships among trust, organizational culture, and perceptions of health care quality and what implications this may or may not have for nursing practice.


Discovery of marker peptides of spirulina microalga proteins for allergen detection in processed foodstuffs.
Spirulina (Arthrospira platensis) proteins were extracted, digested, and analyzed by LC-ESI-FTMS/MS to find highly conserved peptides as markers of the microalga occurrence in foodstuffs. Putative markers were firstly chosen after in silico digestion of allergenic proteins, according to the FAO and WHO criteria, after assuring their presence in food supplements and in (un)processed foodsuffs. Parameters such as sensitivity, sequence size, and uniqueness for spirulina proteins were also evaluated. Three peptides belonging to C-phycocyanin beta subunit (P72508) were designated as qualifiers (ETYLALGTPGSSVAVGVGK and YVTYAVFAGDASVLEDR) and quantifier (ITSNASTIVSNAAR) marker peptides and used to validate the method for linearity, recovery, reproducibility, matrix effects, processing effects, LOD, and LOQ. The main aim was to determine spirulina in commercial foodstuffs like pasta, crackers, and homemade bread incurred with the microalga. The possible inclusion of the designated peptides in a standardized method, based on multiple reaction monitoring using a linear ion trap MS, was also demonstrated.


Effect of adenine nucleotides on cyclooxygenase and lipoxygenase enzyme products of arachidonic acid in human platelets.
Nucleotides are known to enhance cyclooxygenase product formation in several tissues and, in addition, are believed to function as cofactors for mammalian 5-lipoxygenases. Since nucleotides are released by stimulated platelets and by damaged tissue, we examined the hypothesis that nucleotides can affect the metabolism of arachidonic acid (AA) in washed human platelets. The various nucleotides were given 15 sec prior to the addition of 3 microM arachidonic acid and 1 muCi [3H]AA. We found that the phosphorylated adenine derivatives (ATP, ADP, and AMP) increased the formation of 12-hydroxyeicosatetraenoic acid (12-HETE) by 2-fold without altering the formation of cyclooxygenase products. Adenosine was without effect on 12-HETE formation. ATP also stimulated 12-HETE formation in lysed platelets. This suggests that the 12-lipoxygenase enzyme of platelets can be regulated by adenine nucleotides. We next determined the portion of the nucleotide molecule responsible for the enhanced 12-lipoxygenase activity of platelets. Alteration of the nucleotide base led to a decrease in stimulation, with GTP less active than ATP, and UTP even less active than GTP. Studies with adenine nucleotides showed that the length of the phosphate chain was not important. We also found that the stable methylene isosters of ATP (alpha, beta-methylene ATP and beta, gamma-methylene ATP) increased 12-HETE formation, suggesting that the conformation and hydrolysis of the phosphate chain are not responsible for the stimulatory activity. Cyclic 3',5'AMP and 3'AMP were inactive, implying the necessity for a free phosphate at the 5' position for nucleotide stimulation of 12-HETE synthesis. In conclusion, platelet 12-lipoxygenase was stimulated by ATP, as is true for several mammalian 5-lipoxygenases. However, cyclooxygenase product formation by platelets was not altered by nucleotide addition. These studies suggest that following in vivo injury or platelet aggregation, when local concentrations of nucleotides are high, platelet lipoxygenase activity may be stimulated.


Adhesion sutures for seroma reduction in cranial reconstructions with polymethyl methacrylate prosthesis in patients undergoing decompressive craniectomy: A clinical trial.
Cranial reconstruction with polymethyl methacrylate (PMMA) prosthesis is used for calvarial defects secondary to decompressive craniectomies. Seroma is one of the most frequent complications of this procedure and can lead to the dehiscence, extrusion, infection, and loss of the prosthesis. The objective of the study is to analyze the effectiveness of the tacking sutures between the prosthesis and the scalp flap in reducing the seroma. This is a prospective study with 63 patients submitted to cranioplasty between 2014 and 2017 for defects resulting from decompressive craniectomies. All patients were followed up postoperatively for at least 3 months and the diagnosis of seroma was made clinically. In the first 22 patients, the conventional technique was applied and, in the following 41, the technique with tacking sutures was used. The incidence of seroma was collected for both groups. The overall incidence of seroma was 65.1%. Compared to the conventional technique, the use of tacking sutures was associated with a statistically significant reduction in the incidence of seroma from 90.9% to 51.2% (P = 0.002). The use of the tacking sutures in cranioplasties with PMMA prosthesis reduced the incidence of seroma postoperatively.


Causes of unplanned admission after orthopedic procedures in ambulatory surgery.
Orthopaedic procedures performed in Day Surgery Units provide important advantages which disappear when patients require admission when postoperative recovery is not as expected. The aim of this study was to analyse the reasons for unplanned hospital admissions after orthopaedic procedures in a Day Surgery Unit and their relationship between variables such as patient age, anaesthetic risk and technique, procedure or duration. Ambispective cohort study of 5,085 patients who underwent surgical orthopaedic procedures between 1995 and 2017. Thirty-nine variables provided by the Unit's database were analysed. The database was opened on the day of admission and closed the 30th postoperative day. Of the patients, 98.2% were discharged from the Unit. Seventy-four (1.5%) required overnight admission. This percentage showed significant differences in relation to the type of procedure, type of anaesthesia and duration, which conditioned overnight admission due to inadequate postoperative pain management, nausea or wound complications. Seventeen patients (0.3%) required readmission after discharge due to complications that arose at home, such as wound infection, which was the most common. Unplanned admissions are more frequently related to general anaesthesia, lengthy surgeries and procedures such as arthroscopy, hallux valgus corrections or removal of osteosynthesis material. The major reasons for unplanned admissions were inadequate postoperative pain management for overnight admissions and wound infection for admissions after discharge.


Somatic Embryogenesis Induction and Plant Regeneration in Strawberry Tree (Arbutus unedo L.).
Somatic embryogenesis is a powerful tool both for cloning and studies of genetic transformation and embryo development. Most protocols for somatic embryogenesis induction start from zygotic embryos or embryonic-derived tissues which do not allow the propagation of elite trees. In the present study, a reliable protocol for somatic embryogenesis induction from adult trees of strawberry tree is described. Leaves from in vitro proliferating shoots were used to induce somatic embryo formation on a medium containing an auxin and a cytokinin. Somatic embryos germinated in a plant growth regulator-free medium.


Magnetic nanoparticles coated with maltose-functionalized polyethyleneimine for highly efficient enrichment of N-glycopeptides.
Hydrophilic interaction chromatography (HILIC) adsorbents have drawn increasing attention in recent years due to their high efficiency in N-glycopeptides enrichment. The hydrophilicity and binding capacity of HILIC adsorbents are crucial to the enrichment efficiency and mass spectrometry (MS) detection sensitivity of N-glycopeptides. Herein, magnetic nanoparticles coated with maltose-functionalized polyethyleneimine (Fe3O4-PEI-Maltose MNPs) were prepared by one-pot solvothermal reaction coupled with "click chemistry" and utilized for N-glycopeptides enrichment. Owing to the presence of hydrophilic and branched polyethyleneimine, the amount of immobilized disaccharide units was improved about four times. The N-glycopeptides capturing capacity was about 150mg/g (IgG/MNPs) and the MS detection limitation as low as 0.5fmol for IgG and 85% average enrichment recovery were feasibly achieved by using this hybrid magnetic adsorbent. Finally, 1237 unique N-glycosylation sites and 1567 unique N-glycopeptides from 684 N-glycoproteins were reliably characterized from 60μg protein sample extracted from mouse liver. Therefore, this maltose-functionalized polyethyleneimine coated adsorbent can play a promising role in highly efficient N-glycopeptides enrichment for glycoproteomic analyses of complex protein samples.


The impact of developing a pectus center for chest wall deformities.
In 2011, we established a dedicated center for patients with chest wall deformities. Here, we evaluate the center's effect on patient volume and management. A retrospective review of 699 patients with chest wall anomalies was performed. Patients were compared, based on the date of initial consultation, before the pectus center opened (July 2009-June 2011, Group 1) versus after (July 2011-June 2013, Group 2). Analysis was performed utilizing Chi-square and Mann-Whitney U tests. 320 patients were in Group 1 and 379 in Group 2, an 18.4 % increase in patient volume. Excavatum patients increased from 172 (Group 1) to 189 (Group 2). Carinatum patients increased from 125 (Group 1) to 165 (Group 2). Patients undergoing operative repair of carinatum/mixed defects dropped significantly from 15 % (Group 1) to 1 % (Group 2) (p < 0.01), whereas those undergoing nonoperative bracing for carinatum/mixed defects rose significantly from 19 % (Group 1) to 63 % (Group 2) (p < 0.01). Patients traveled 3-1249 miles for a single visit. Initiating a dedicated pectus center increased patient volume and provided an effective transition to nonoperative bracing for carinatum patients. The concentrated focus of medical staff dedicated to chest wall deformities has allowed us to treat patients on a local and regional level.


Calcitonin COOH-terminal cleavage peptide as a model for identification of novel neuropeptides predicted by recombinant DNA analysis.
A strategy is presented for identification of putative neurohormones that were predicted by sequence analysis of recombinant cDNa molecules. The nucleotide sequence of a cloned calcitonin cDNA insert predicts that the proteolytic excision of calcitonin from a 136-amino acid precursor should generate two additional peptides derived from regions flanking the calcitonin sequence on the NH2 and carboxyl termini. The predicted 16-amino acid COOH-terminal cleavage peptide (NH2-Asp-Met-Ala-Lys-Asp-Leu-Glu-Thr-Asn-His-His-Pro-Thr-Phe-Gly-Asn-COOH) was chemically synthesized using solid phase procedures. Antibodies against this synthetic hexadecapeptide detect immunologically cross-reactive material in tissue extracts from normal thyroid glands and calcitonin-producing medullary thyroid carcinomas. Each of the four calcitonin mRNA-directed cell-free translation products contain the immunological determinants for both calcitonin and the COOH-terminal cleavage peptide.


Cross-linking of glutathione to DNA by cancer chemotherapeutic platinum coordination complexes.
Glutathione can modulate the toxicity of a variety of drugs, although its role in modulating toxicity by anticancer platinum drugs is ambivalent. At physiologically relevant concentrations, glutathione can inhibit the reaction between DNA and cis-dichloro(ethylenediamine)platinum(II) (cis-DEP). Glutathione can also react with monofunctional adducts in DNA to produce a glutathione-Pt-deoxyguanosine cross-link which would reduce the potential toxicity of the drug. The relative importance of these two mechanisms of detoxification is unknown, although both mechanisms probably contribute to glutathione modulation of platinum toxicity.


Isolation and characterization of some antioxidative compounds from the rhizomes of smaller galanga (Alpinia officinarum Hance).
Antioxidative compounds were isolated from the methanol extract of fresh rhizome of smaller galanga (Alpinia officinarum Hance). Seven phenylpropanoids (1-7) were finally obtained by reversed-phase HPLC, and their structures were elucidated by MS and NMR analyses. They comprised the two known compounds, (E)-p-coumaryl alcohol gamma-O-methyl ether (1) and (E)-p-coumaryl alcohol (6), and the five novel compounds, stereoisomers of (4E)-1,5-bis(4-hydroxyphenyl)-1-methoxy-2-(methoxymethyl)-4-pentene (2a and 2b), stereoisomers of (4E)-1,5-bis(4-hydroxyphenyl)-1-ethoxy-2-(methoxymethyl)-4-pentene (3a and 3b), (4E)-1,5-bis(4-hydroxyphenyl)-1-[(2E)-3-(4-acetoxyphenyl)-2-propenoxy]-2-(methoxymethyl)-4-pentene (4), (4E)-1,5-bis(4-hydroxyphenyl)-2-(methoxymethyl)-4-penten-1-ol (5), and (4E)-1,5-bis(4-hydroxyphenyl)-2-(hydroxymethyl)-4-penten-1-ol (7). Compounds 1-7 were detected for the first time as constituents of galanga rhizomes and exhibited antioxidative activities against the autoxidation of methyl linoleate in bulk phase.


Using quantitative trait in adults with ADHD to test predictions of dual-process theory.
Dual-process theory is a widely utilized modelling tool in the behavioral sciences. It conceptualizes decision-making as an interaction between two types of cognitive processes, some of them fast and intuitive, others slow and reflective. We make a novel contribution to this literature by exploring differences between adults with clinically diagnosed ADHD and healthy controls for a wide range of behaviors. Given the clinical picture and nature of ADHD symptoms, we had a strong a priori reason to expect differences in intuitive vs reflective processing; and thus an unusually strong case for testing the predictions of dual-process theory. We found mixed results, with overall weaker effects than expected, except for risk taking, where individuals with ADHD showed increased domain sensitivity for gains vs losses. Some of our predictions were supported by the data but other patterns are more difficult to reconcile with theory. On balance, our results provide only limited empirical support for using dual-process theory to understand basic social and economic decision-making.


Bactericidal action mechanism of negatively charged food grade clove oil nanoemulsions.
Clove oil (CO) anionic nanoemulsions were prepared with varying ratios of CO to canola oil (CA), emulsified and stabilized with purity gum ultra (PGU), a newly developed succinylated waxy maize starch. Interfacial tension measurements showed that CO acted as a co-surfactant and there was a gradual decrease in interfacial tension which favored the formation of small droplet sizes on homogenization until a critical limit (5:5% v/v CO:CA) was reached. Antimicrobial activity of the negatively charged CO nanoemulsion was determined against Gram positive GPB (Listeria monocytogenes and Staphylococcus aureus) and Gram negative GNB (Escherichia coli) bacterial strains using minimum inhibitory concentration (MIC) and a time kill dynamic method. Negatively charged PGU emulsified CO nanoemulsion showed prolonged antibacterial activities against Gram positive bacterial strains. We concluded that negatively charged CO nanoemulsion droplets self-assemble with GPB cell membrane, and facilitated interaction with cellular components of bacteria. Moreover, no electrostatic interaction existed between negatively charged droplets and the GPB membrane.


NAVIGATE 24-Month Results: Electromagnetic Navigation Bronchoscopy for Pulmonary Lesions at 37 Centers in Europe and the United States.
Electromagnetic navigation bronchoscopy (ENB) is a minimally invasive, image-guided approach to access lung lesions for biopsy or localization for treatment. However, no studies have reported prospective 24-month follow-up from a large, multinational, generalizable cohort. This study evaluated ENB safety, diagnostic yield, and usage patterns in an unrestricted, real-world observational design. The NAVIGATE single-arm, pragmatic cohort study (NCT02410837) enrolled subjects at 37 academic and community sites in seven countries with prospective 24-month follow-up. Subjects underwent ENB using the superDimension navigation system versions 6.3 to 7.1. The prespecified primary end point was procedure-related pneumothorax requiring intervention or hospitalization. A total of 1388 subjects were enrolled for lung lesion biopsy (1329; 95.7%), fiducial marker placement (272; 19.6%), dye marking (23; 1.7%), or lymph node biopsy (36; 2.6%). Concurrent endobronchial ultrasound-guided staging occurred in 456 subjects. General anesthesia (78.2% overall, 56.6% Europe, 81.4% United States), radial endobronchial ultrasound (50.6%, 4.0%, 57.4%), fluoroscopy (85.0%, 41.7%, 91.0%), and rapid on-site evaluation use (61.7%, 17.3%, 68.5%) differed between regions. Pneumothorax and bronchopulmonary hemorrhage occurred in 4.7% and 2.7% of subjects, respectively (3.2% [primary end point] and 1.7% requiring intervention or hospitalization). Respiratory failure occurred in 0.6%. The diagnostic yield was 67.8% (range: 61.9%-70.7%; 55.2% Europe, 69.8% United States). Sensitivity for malignancy was 62.6%. Lung cancer clinical stage was I to II in 64.7% (55.3% Europe, 65.8% United States). Despite a heterogeneous cohort and regional differences in procedural techniques, ENB demonstrates low complications and a 67.8% diagnostic yield while allowing biopsy, staging, fiducial placement, and dye marking in a single procedure.


A new enzymic method for the isolation and culture of human bladder body smooth muscle cells.
Cultured cells of the human urinary bladder smooth muscle are useful for investigating bladder function, but methods for culturing them are not well developed. We have now established a novel enzymic technique. The smooth muscle layer was separated out and incubated with 0.2% trypsin for 30 min at 37 degrees C. The samples were then minced and incubated with 0.1% collagenase for 30 min and centrifuged at 900 g. The pellets were resuspended in RPMI-1640 medium containing 10% fetal calf serum (FCS) and centrifuged at 250 g. The smooth muscle cells from the supernatant were cultured in RPMI-1640 containing 10% FCS. The cells grew to confluence after 7-10 days, forming the "hills and valleys" growth pattern characteristic of smooth muscle cells. Immunostaining with anti-alpha-actin, anti-myosin, and anti-caldesmon antibodies demonstrated that 99% of the cells were smooth muscle cells. To investigate the pharmacological properties of the cultured cells, we determined the inhibitory effect of muscarinic receptor antagonists on the binding of [3H]N-methylscopolamine to membranes from cultured cells. The pKi values obtained for six antagonists agreed with the corresponding values for transfected cells expressing the human muscarinic M2 subtype. Furthermore, carbachol produced an increase in the concentration of cytoplasmic free Ca2+ an action that was blocked by 4-diphenylacetoxy-N-methylpiperidine methiodide, an M3 selective antagonist. This result suggests that these cells express functional M3 muscarinic receptors, in addition to M2 receptors. The subcultured cells therefore appear to be unaffected by our new isolation method.


Interferon-induced protein Mx accumulates in nuclei of mouse cells expressing resistance to influenza viruses.
In mouse cells carrying the dominant influenza resistance allele Mx+ (but not in Mx- cells) interferon-alpha/beta (IFN) induces an efficient antiviral state against influenza viruses and, concomitantly, the synthesis of a 75,000-Da protein (protein Mx). Here, indirect immunofluorescence using monoclonal antibodies was used to demonstrate that protein Mx accumulates in the nucleus of IFN-treated Mx+ cells, suggesting a nuclear site of action. Protein Mx is present in the nucleus of untreated influenza virus-resistant macrophages freshly explanted from the peritoneal cavity of Mx+ mice but is lost with time in culture when peritoneal macrophages become permissive for influenza virus.


Depth profiling with modified dc-Grimm and rf-Grimm-type glow discharges operated with high gas flow rates and coupled to a high-resolution mass spectrometer.
The improved analytical capability of direct-current (dc) and radiofrequency (rf) "fast flow" glow discharges coupled to a sector field mass spectrometer (GD-SFMS) are presented. In particular, the effect of GD chamber design has been studied to obtain suitable crater shapes for depth-profile analysis of solid samples while maintaining the high sensitivity and stability of this source. In this study it was observed that the distance between the sample surface and the end of the flow tube is critical and so careful optimisation is needed. Under optimum conditions plane crater profiles, with high ion-signal sensitivity and sufficient stability, were obtained. The capability to determine qualitative and semi-quantitative depth profiles is presented here using, as model, a coated sample of certified thickness. Finally, the depth resolution achieved for qualitative depth profiles obtained by rf-GD-(SF)MS is compared with that for the well-established rf-GD optical emission spectroscopy (OES) technique.


Marital break-up among couples raising families by artificial insemination by donor.
The separation rate for Norwegian women who had given birth to children after artificial insemination by donor (AID, n = 227) was compared with the separation rate among all Norwegian women that met certain demographic criteria for equality (controls, n = 6,689). The criteria included identical year of maternal birth, similar age at marriage, and equally long period between marriage and birth of the first child. Mean duration of marriage at first delivery was 7.4 years, and mean age of the first child was 3.6 years at registration of the data. The incidence of marital break-up in the AID group was 1.8%, as compared with 3.0% to 3.7% in the control group. The difference is not significant (P approximately 0.15). The authors conclude that the separation rate in AID families does not differ significantly from the observed separation rate in a demographically matched population. The study highlights the importance of selecting a proper control group before statements of the magnitude of separations in AID families are made.


Patterns and mutational signatures of tandem base substitutions causing human inherited disease.
Tandem base substitutions (TBSs) are multiple mutations that comprise two or more contiguous nucleotide substitutions without any net gain or loss of bases. They have recently become recognized as a distinct category of human genomic variant. However, their role in causing human inherited disease so far has not been studied methodically. Here, using data from the Human Gene Mutation Database (http://www.hgmd.org), we identified 477 events to be TBSs (doublets, 448; triplets, 16; and quadruplets to octuplets, 13). A comprehensive sequence pattern and context analysis implied the likely fundamental importance of translesion synthesis (TLS) DNA polymerases in generating these diverse TBSs but revealed that TLS polymerases may operate differently in generating TBSs of ≤ 3 bases (bypass of endogenous DNA lesions) than those of ≥ 4 bases (serial replication slippage). Moreover, GC was found to be the most frequently affected dinucleotide with GC/GC>AA/TT being the most frequent double TBS. Comparison with cancer genome mutational spectra allowed us to conclude that human germline TBSs arise predominantly through the action of endogenous mechanisms of mutagenesis rather than through exposure to exogenous mutagens. Finally, the rates of double and triple TBSs were estimated to be 0.2-1.2 × 10(-10) and 0.8-4.8 × 10(-12) per base per generation, respectively.


Awareness and uptake of measures for preventing CNS birth defects among mothers of affected children in a sub-Saharan African neurosurgeon's practice.
The two main measures used in the reduction of the birth prevalence of CNS birth defects are (i) folic acid, FA, supplementation/food fortification for reproductively active women as the primary one, and (ii) timely screening/intrauterine diagnosis, and possibly termination of the affected pregnancies. We performed a cross-sectional survey of the levels of awareness and uptake of these measures in a consecutive cohort of mothers of children with CNS birth defects presenting for neurosurgical treatment in an African clinical practice. There were 151 cases, 101 of them neural tube defects, NTDs. The level of awareness of the role of FA in prevention of these defects was low, 18.8%, and dietary multivitamin supplementation was ingested by only10.7% of the study subjects. The mothers' obstetric behavior in the index pregnancies was suboptimal: pregnancy registration and commencement of obstetric supplements were at median gestational age of 4 months, and obstetric ultrasonography was obtained infrequently, and in an unregulated milieu. Only 17.8% of these CNS birth defects were diagnosed prenatally, but >80% of the mothers would have liked to have the intrauterine diagnosis, and about 23% might have asked for termination of these pregnancies. The levels of awareness and uptake of measures for preventing CNS birth defects among mothers of affected children in this sub-Saharan African women cohort are low. Interestingly, many of the mothers were very favorably disposed to receiving, and acting on, the information about the screen detected CNS birth defects in their fetuses.


Integrating the Xpert MTB/RIF assay into a diagnostic workflow for rapid detection of Mycobacterium tuberculosis in a low-prevalence area.
The Xpert MTB/RIF assay is a rapid and fully automated real-time PCR assay. The performance of the Xpert MTB/RIF assay as a primary screening test for urgent clinical specimens was evaluated during a 2-year period. The results showed that replacing smear microscopy with the Xpert MTB/RIF assay facilitates laboratory handling and improves the sensitivity and specificity of Mycobacterium tuberculosis detection.


An Overview of Flow Cytometry: Its Principles and Applications in Allergic Disease Research.
Flow cytometry is a popular technique used for both clinical and research purposes. It involves laser-based technology to characterize cells based on size, shape, and complexity. Additionally, flow cytometers are equipped with the ability to take fluorescence measurements at multiple wavelengths. This capability makes the flow cytometer a practical resource in the utilization of fluorescently conjugated antibodies, fluorescent proteins, DNA binding dyes, viability dyes, and ion indicator dyes. As the technology advances, the number of parameters a flow cytometer can measure has increased tremendously, and now some has the capacity to analyze 30-50 or more parameters on a single cell. Here, we describe the basic principles involved in the mechanics and procedures of flow cytometry along with an insight into applications of flow cytometry techniques for biomedical and allergic disease research.


Disrupted minor intron splicing is prevalent in Mendelian disorders.
Splicing is crucial for proper gene expression, and is predominately executed by the major spliceosome. Conversely, 722 introns in 699 human minor intron-containing genes (MIGs) are spliced by the minor spliceosome. Splicing of these minor introns is disrupted in diseases caused by pathogenic variants in the minor spliceosome, ultimately leading to the aberrant expression of a subset of these MIGs. However, the effect of variants in minor introns and MIGs on diseases remains unexplored. Variants in MIGs and associated clinical manifestations were identified using ClinVar. The HPO database was then used to curate the related symptoms and affected organ systems. We found pathogenic variants in 211 MIGs, which commonly resulted in intellectual disability, seizures and microcephaly. This revealed a subset of MIGs whose aberrant splicing may contribute to the pathogenesis of minor spliceosome-related diseases. Moreover, we identified 51 pathogenic variants in minor intron splice sites that reduce the splice site strength and can induce alternative splicing. These findings highlight that disrupted minor intron splicing has a broader impact on human diseases than previously appreciated. The hope is that this knowledge will aid in the development of therapeutic strategies that incorporate the minor intron splicing pathway.


Subcutaneous microvascular (capillary) calcification. Another basis for livedo-like skin changes?
A morbidly obese woman with acute renal failure, high serum phosphorus and slightly depressed serum calcium levels, developed areas of induration in the subcutis with associated livedo reticularis. Later, the subcutis became necrotic and the skin ulcerated. The arterioles, and notably widened occluded capillaries, were found to contain calcium and phosphorus as determined by energy dispersive spectrometry. X-ray diffraction of the subcutis in an early stage of this lesion showed that the mineral was most likely a poorly crystalline hydroxyapatite.


Thermal unfolding molecular dynamics simulation of Escherichia coli dihydrofolate reductase: thermal stability of protein domains and unfolding pathway.
Temperature induced unfolding of Escherichia coli dihydrofolate reductase was carried out by using molecular dynamic simulations. The simulations show that the unfolding generally involves an initial end-to-end collapse of the adenine binding domain into partially extended loops, followed by a gradual breakdown of the remaining beta sheet core structure. The core, which consists of beta strands 5-7, was observed to be the most resistant to thermal unfolding. This region, which is made up of part of the N terminus domain and part of the large domain of the E. coli dihydrofolate reductase, may constitute the nucleation site for protein folding and may be important for the eventual formation of both domains. The unfolding of different domains at different stages of the unfolding process suggests that protein domains vary in stability and that the rate at which they unfold can affect the overall outcome of the unfolding pathway. This observation is compared with the recently proposed hierarchical folding model. Finally, the results of the simulation were found to be consistent with a previous experimental study (Frieden, Proc Natl Acad Sci USA 1990;87:4413-4416) which showed that the folding process of E. coli dihydrofolate reductase involves sequential formation of the substrate binding sites.


Insight into the changing patterns in clinical and academic activities of the orthopedic residents during COVID-19 pandemic: a cross-sectional survey.
The COVID-19 pandemic has presented an unprecedented challenge to the orthopedic residency training programs to adapt to a form of a web-based learning process and simulation-based training. This study focusses on the viewpoint of the orthopedic residents to the paradigm shift in clinical care as well as the academic activities. An anonymous questionnaire was created in an online survey generator and was sent through e-mail to 227 orthopedic residents of seven tertiary care centres in North India. The questionnaire was divided into three sections, academic activity section, mental health section, and clinical activity section. There were a total of 44 single answer questions with answers according to the increasing difficulty at present situation compared to a previous time before the COVID-19 pandemic. A total of 158 questionnaires were filled by 107 junior residents (67.7%) and 51 senior residents (32.3%). 49 residents (31%) were quarantined and three became positive for COVID-19. Although all of them knew about necessary precautions, personal protective equipment was difficult to avail at times. Increased difficulty in recruiting new patients for research (48.9%) and conducting prospective research (48.7%) was observed. The online-based learning process was reported to be easier (44.2%) by most of the residents. Routine clinical work in the operating room, out-patient department, and inpatient department was found to be difficult according to the majority of the residents along with the anxiety of contracting the infection. There are unique opportunities for improvement of residency programs during these times of uncertainty and the findings of this study can help the universities as well as program chairs to develop a robust program that can outlive this pandemic. The web-based learning process might prove to be useful and can be incorporated into the resident training program in the long term. Level V.


Photodegradation of Rhodamine B using gallium hybrids as an efficient photocatalyst.
Keeping in view the toxicity of the Rhodamine B, the present study is designed to remediate the water loaded with toxic dyes using gallium oxide and gallium hybrids as photocatalyst. Precipitation coupled with sonochemical method is adopted for the synthesis of gallium oxide while the post grafting method is adopted for the synthesis of gallium hybrids with the indole and its derivatives. FTIR spectra showed the characteristic absorption bands of gallium oxide and gallium hybrids at 400-700 cm-1 and 1400-1600 cm-1. SEM and XRD showed the micro-sized rectangular rod-shaped gallium oxide with rhombohedral geometry. The average crystallite size of gallium hybrids was 26-32 nm calculated using the Debye Scherrer and Williamson-Hal models. The BET isotherm of gallium hybrids revealed the adsorption type-IV and hysteresis loop (H3) proposing multilayer and mesoporous structures with increase in surface area from 26 m2/g of gallium oxide to 31 m2/g of gallium-indole, 35 m2/g of gallium-methyl indole, and 37 m2/g of gallium-carboxylic indole. XPS showed the presence of gallium (3-14%), oxygen (28-32%), nitrogen (23-46%), and carbon (9-46%). The gallium oxide and gallium hybrids showed 47-72% optimum degradation of Rhodamine B under 2 h of illumination at pH 7 and 0.03 mg/L. The degradation rate followed a Langmuir-Hinshelwood model with R2 > 0.9.


Increased insufflation pressure enhances the development of liver metastasis in a mouse laparoscopy model: possible mechanisms.
The effect of different insufflation pressures and durations of CO2 pneumoperitoneum on the growth of liver metastasis was investigated in a mouse model. The possible mechanisms involved in the pressure-related enhancement of liver metastasis were also examined. Mice inoculated intraportally with colon 26 cells underwent CO2 pneumoperitoneum at different pressures (5,10, or 15 mmHg) for 30 or 60 min, or received no treatment other than tumor cell inoculation (control). The subsequent growth of liver metastases was examined. Mice injected intraportally with 111In-oxine-labeled colon 26 cells underwent pneumoperitoneum at three different pressures or served as controls. The radioactivity of the liver was determined to evaluate tumor accumulation in the liver. Mice received pneumoperitoneum at three different pressures or received trocar placement alone. Changes in plasma interleukin-6 levels were determined. The growth of liver metastases on day 14 was influenced by increased insufflation pressures (p < 0.05) rather than the prolonged duration of pneumoperitoneum without significant interaction. The 15-mmHg pneumoperitoneum group showed a higher (p < 0.05) accumulation of radioactivity in the liver compared with the 5-mmHg pneumoperitoneum group and controls. Pneumoperitoneum groups with 5 and 10 mmHg showed higher (p < 0.05) peak levels of IL-6 compared with controls. An elevated insufflation pressure plays an important role in the enhancement of liver metastases, and this pressure-related adverse effect may be partly relevant to facilitating accumulation of tumor cells in the liver.


Regulation of membrane dynamics in developing epithelia.
A cellular understanding of developmental mechanisms embraces complex dynamic processes at the cell surface. Studies of trafficking pathways and of associated cytoskeletal elements in genetically tractable organisms allow a better analysis of the mechanisms underlying the control of cell polarization and tissue morphogenesis at different stages of animal development. It is now possible to identify signalling pathways that provide the temporal triggers and/or spatial cues to regulate core cellular effectors during migration, morphogenesis and the formation of epithelia.


'In silico expression analysis', a novel PathoPlant web tool to identify abiotic and biotic stress conditions associated with specific cis-regulatory sequences.
Using bioinformatics, putative cis-regulatory sequences can be easily identified using pattern recognition programs on promoters of specific gene sets. The abundance of predicted cis-sequences is a major challenge to associate these sequences with a possible function in gene expression regulation. To identify a possible function of the predicted cis-sequences, a novel web tool designated 'in silico expression analysis' was developed that correlates submitted cis-sequences with gene expression data from Arabidopsis thaliana. The web tool identifies the A. thaliana genes harbouring the sequence in a defined promoter region and compares the expression of these genes with microarray data. The result is a hierarchy of abiotic and biotic stress conditions to which these genes are most likely responsive. When testing the performance of the web tool, known cis-regulatory sequences were submitted to the 'in silico expression analysis' resulting in the correct identification of the associated stress conditions. When using a recently identified novel elicitor-responsive sequence, a WT-box (CGACTTTT), the 'in silico expression analysis' predicts that genes harbouring this sequence in their promoter are most likely Botrytis cinerea induced. Consistent with this prediction, the strongest induction of a reporter gene harbouring this sequence in the promoter is observed with B. cinerea in transgenic A. thaliana. DATABASE URL: http://www.pathoplant.de/expression_analysis.php.


Lens morphometry determined by B-mode ultrasonography of the normal and cataractous canine lens.
To determine axial lens thickness, anterior chamber depth and axial globe length in canine eyes with normal lenses and in eyes with immature, mature, congenital, posterior polar and diabetic cataract. B-mode ultrasonography was performed in 50 normal dogs and, as a prephacoemulsification screening procedure, in 100 dogs with cataract. Axial B-mode ultrasonograms were used to determine lens thickness, anterior chamber depth and globe diameter. Statistical comparisons between groups were made by analysis of variance and multivariate analysis, with a significance level of P < 0.05. Axial globe lengths were not statistically significantly different between groups apart from the smaller globes in younger dogs with congenital cataract. Axial lens thickness in diabetics (8.4 +/- 0.9 mm) was statistically significantly different from the lens thickness in normal eyes (6.7 +/- 1.0 mm), eyes with immature cataract (6.4 +/- 0.8 mm) and eyes with mature cataract (7.4 +/- 0.9 mm) although these groups, while varying in thickness, were not statistically significantly different from each other. Anterior chamber depth was statistically significantly reduced in eyes with diabetic cataract (2.9 +/- 0.1 mm) from that in normal eyes (3.8 +/- 0.1 mm), eyes with immature cataract (3.5 +/- 0.1 mm) and eyes with mature cataract (3.2 +/- 0.6 mm) although these groups, while varying in chamber depth, were not statistically significantly different from each other. Lenses with diabetic cataracts were significantly increased in axial thickness compared to lenses in other eyes, although lenses with mature cataracts showed a trend towards increased axial thickness and immature cataracts demonstrated a trend towards reduced thickness. While previous studies on cataract pathobiology have suggested a reduction in lens thickness in immature cataract through lens protein loss and an increase in thickness in mature cataracts through intumescence, this study is the first to document these changes in the canine lens.


Retrospective cost-effectiveness of the 23-valent pneumococcal polysaccharide vaccination program in Australia.
The Australian infant pneumococcal vaccination program was funded in 2005 using the 7-valent pneumococcal conjugate vaccine (PCV7) and the 13-valent conjugate vaccine (PCV13) in 2011. The PCV7 and PCV13 programs resulted in herd immunity effects across all age-groups, including older adults. Coincident with the introduction of the PCV7 program in 2005, 23-valent pneumococcal polysaccharide vaccine (PPV23) was funded for all Australian adults aged over 65 years. A multi-cohort Markov model with a cycle length of one year was developed to retrospectively evaluate the cost-effectiveness of the PPV23 immunisation program from 2005 to 2015. The analysis was performed from the healthcare system perspective with costs and quality-adjusted life years discounted at 5% annually. The incremental cost-effectiveness ratio (ICER) for PPV23 doses provided from 2005 to 2015 was calculated separately for each year when compared to no vaccination. Parameter uncertainty was explored using deterministic and probabilistic sensitivity analysis. It was estimated that PPV23 doses given out over the 11-year period from 2005 to 2015 prevented 771 hospitalisations and 99 deaths from invasive pneumococcal disease (IPD). However, the estimated IPD cases and deaths prevented by PPV23 declined by more than 50% over this period (e.g. from 12.9 deaths for doses given out in 2005 to 6.1 in 2015), likely driven by herd effects from infant PCV programs. The estimated ICER over the period 2005 to 2015 was approximately A$224,000/QALY gained compared to no vaccination. When examined per year, the ICER for each individual year worsened from $140,000/QALY in 2005 to $238,000/QALY in 2011 to $286,000/QALY in 2015. The cost-effectiveness of the PPV23 program in older Australians was estimated to have worsened over time. It is unlikely to have been cost-effective, unless PPV23 provided protection against non-invasive pneumococcal pneumonia and/or a low vaccine price was negotiated. A key policy priority should be to review of the future use of PPV23 in Australia, which is likely to be more cost-effective in certain high-risk groups.


Tyrosine as important contributor to the antioxidant capacity of seminal plasma.
A novel post-addition method, based on the trapping of ABTS-radicals, is applied for studying the total antioxidant capacity of seminal plasma. A remarkable profile is observed, in which seminal plasma quenches radicals in a continuous, relatively slow fashion. Five putative antioxidants present in seminal plasma were studied using the same assay. Some of the compounds such as ascorbic acid, alpha-tocopherol and uric acid exert immediate, fast radical trapping, whereas hypotaurine and tyrosine give rise to the same slow radical trapping curve as seminal plasma. Due to this slow, continuous radical trapping, quantification of the total antioxidant capacity (expressed as trolox equivalent antioxidant capacity, TEAC) strongly depends on the chosen time point after onset of radical trapping. When determined during the slow antioxidant trapping phase, tyrosine has a powerful antioxidant capacity, which in combination with its relatively high plasma concentration makes it an important contributor to the total antioxidant capacity of seminal plasma.


Thiol proteases. Comparative studies based on the high-resolution structures of papain and actinidin, and on amino acid sequence information for cathepsins B and H, and stem bromelain.
An accurate three-dimensional structure is known for papain (1.65 A resolution) and actinidin (1.7 A). A detailed comparison of these two structures was performed to determine the effect of amino acid changes on the conformation. It appeared that, despite only 48% identity in their amino acid sequence, different crystallization conditions and different X-ray data collection techniques, their structures are surprisingly similar with a root-mean-square difference of 0.40 A between 76% of the main-chain atoms (differences less than 3 sigma). Insertions and deletions cause larger differences but they alter the conformation over a very limited range of two to three residues only. Conformations of identical side-chains are generally retained to the same extent as the main-chain conformation. If they do change, this is due to a modified local environment. Several examples are described. Spatial positions of hydrogen bonds are conserved to a greater extent than are the specific groups involved. The greatest structural similarity is found for the active site residues of papain and actinidin, for the internal water molecules and for the main-chain conformation of residues in alpha-helices and anti-parallel beta-sheet structure. This was reflected also in the similarity of the temperature factors. It suggests that the secondary structural elements form the skeleton of the molecule and that their interaction is the main factor in directing the fold of the polypeptide chain. Therefore, substitution of residues in the skeleton will, in general, have the most drastic effect on the conformation of the protein molecule. In papain and actinidin, some main-chain-side-chain hydrogen bonds are also strongly conserved and these may determine the folding of non-repetitive parts of the structure. Furthermore, we included primary structure information for three homologous thiol proteases: stem bromelain, and the cathepsins B and H. By combining the three-dimensional structural information for papain and actinidin with sequence homologies and identities, we conclude that the overall folding pattern of the polypeptide chain is grossly the same in all five proteases, and that they utilize the same catalytic mechanism.


Histologic evaluation of chronic human wounds treated with hydrocolloid and nonhydrocolloid dressings.
Studies of acute surgical wounds in pigs have shown granulomatous tissue reactions induced by hydrocolloid dressings. We examined biopsy specimens from chronic wounds in humans treated with hydrocolloid or nonhydrocolloid dressings to determine whether such reactions occurred. A blinded retrospective analysis of 22 chronic ulcer biopsy specimens from 22 patients was performed by two dermatopathologists independently. No differences were seen between hydrocolloid-treated and non-hydrocolloid-treated wounds. Granulation tissue was seen in all specimens. No cavities, eosinophilic inclusions, polarizable material, or multinucleated giant cells were identified. There are no significant histologic differences between hydrocolloid-treated and non-hydrocolloid-treated wounds in humans.


[Clinical and immunological parallels in soft tissue tumors].
The authors report the results of studying the cell immunity by skin delayed hypersensitivity response to DNCB and tuberculin as well as the reactions of spontaneous rosette-formation, blasttransformation to PHA, inhibition of leucocyte migration in 88 patients with soft tissue sarcomas, in 12 patients with desmoids, the control group comprised 19 patients with benign soft tissue tumors. The study has indicated a reliably impaired cell immunity in patients with sarcomas.


Direct observation of the evolution of cell-type-specific microRNA expression signatures supports the hematopoietic origin model of endothelial cells.
The evolution of specialized cell-types is a long-standing interest of biologists, but given the deep time-scales very difficult to reconstruct or observe. microRNAs have been linked to the evolution of cellular complexity and may inform on specialization. The endothelium is a vertebrate-specific specialization of the circulatory system that enabled a critical new level of vasoregulation. The evolutionary origin of these endothelial cells is unclear. We hypothesized that Mir-126, an endothelial cell-specific microRNA may be informative. We here reconstruct the evolutionary history of Mir-126. Mir-126 likely appeared in the last common ancestor of vertebrates and tunicates, which was a species without an endothelium, within an intron of the evolutionary much older EGF Like Domain Multiple (Egfl) locus. Mir-126 has a complex evolutionary history due to duplications and losses of both the host gene and the microRNA. Taking advantage of the strong evolutionary conservation of the microRNA among Olfactores, and using RNA in situ hybridization, we localized Mir-126 in the tunicate Ciona robusta. We found exclusive expression of the mature Mir-126 in granular amebocytes, supporting a long-proposed scenario that endothelial cells arose from hemoblasts, a type of proto-endothelial amoebocyte found throughout invertebrates. This observed change of expression of Mir-126 from proto-endothelial amoebocytes in the tunicate to endothelial cells in vertebrates is the first direct observation of the evolution of a cell-type in relation to microRNA expression indicating that microRNAs can be a prerequisite of cell-type evolution.


The political economy of a public health case management program's transition into medical homes.
Throughout the United States, public health leaders are experimenting with how best to integrate services for individuals with complex needs. To that end, North Carolina implemented a policy incorporating both local public health departments and other providers into medical homes for low income pregnant women and young children at risk of developmental delays. To understand how this transition occurred within local communities, a pre-post comparative case study was conducted. A total of 42 people in four local health departments across the state were interviewed immediately before the 2011 policy change and six months later: 32 professionals (24 twice) and 10 pregnant women receiving case management at the time of the policy implementation. We used constant comparative analysis of interview and supplemental data to identify three key consequences of the policy implementation. One, having medical homes increased the centrality of other providers relative to local health departments. Two, a shift from focusing on personal relationships toward medical efficiency diverged in some respects from both case managers' and mothers' goals. Three, health department staff re-interpreted state policies to fit their public health values. Using a political economy perspective, these changes are interpreted as reflecting shifts in public health's broader ideological environment. To a large extent, the state successfully induced more connection between health department-based case managers and external providers. However, limited provider engagement may constrain the implementation of the envisioned medical homes. The increased focus on medical risk may also undermine health departments' role in supporting health over time by attenuating staff relationships with mothers. This study helps clarify how state public health policy innovations unfold at local levels, and why front line practice may in some respects diverge from policy intent.


Adrenergic and cholinergic innervation of the mammary gland in the pig.
Adrenergic and acetylcholinesterase-positive (AChE-positive) innervation of the mammary gland in the sexually immature and mature pigs was studied using histochemical methods. Upon examining the adrenergic and cholinergic innervation, the adrenergic innervation was found to be much more developed. The majority of both sub-populations of the nerve fibres studied was localized in the subcutaneous tissue of the mammary gland. Adrenergic and AChE-positive nerve fibres also supplied structures of the nipple (subcutaneous tissue, blood vessels, smooth muscles fibres) and glandular tissue (blood vessels, lactiferous ducts). The glandular tissue contained the smallest number of adrenergic and AChE-positive nerve fibres. No distinct differences were observed in the adrenergic and AChE-positive innervation of the porcine mammary gland between the juvenile and non-pregnant adult animals.


A randomized controlled trial of two primary school intervention strategies to prevent early onset tobacco smoking.
In this article, we examine the impact of two universal, grade 1 preventive interventions on the onset of tobacco smoking as assessed in early adolescence. The classroom-centered (CC) intervention was designed to reduce the risk for tobacco smoking by enhancing teachers' behavior management skills in first grade and, thereby, reducing child attention problems and aggressive and shy behavior-known risk behaviors for later substance use. The family-school partnership (FSP) intervention targeted these early risk behaviors via improvements in parent-teacher communication and parents' child behavior management strategies. A cohort of 678 urban, predominately African-American, public school students were randomly assigned to one of three Grade 1 classrooms at entrance to primary school (age 6). One classroom featured the CC intervention, a second the FSP intervention, and the third served as a control classroom. Six years later, 81% of the students completed audio computer-assisted self-interviews. Relative to controls, a modest attenuation in the risk of smoking initiation was found for students who had been assigned to either the CC or FSP intervention classrooms (26% versus 33%) (adjusted relative risk for CC/control contrast=0.57, 95% confidence interval (CI), 0.34-0.96; adjusted relative risk for FSP/control contrast=0.69, 95% CI, 0.50-0.97). Results lend support to targeting the early antecedent risk behaviors for tobacco smoking.