Factors affecting stability of z-ligustilide in the volatile oil of radix angelicae sinensis and ligusticum chuanxiong and its stability prediction.
The purpose of this investigation is to obtain a suitable vehicle for Z-ligustilide in the volatile oil of Radix Angelicae Sinensis and Ligusticum Chuanxiong in which it is stable enough for the application in pharmaceutics, to investigate its degradation laws, and to predict its shelf-life at 25 degrees C. Factors including temperature, light, pH value, co-solvents and antioxidants can all influence the stability of Z-ligustilide, thereinto antioxidants could markedly improve its stability in aqueous solution by almost 35%. The suitable vehicle for Z-ligustilide contains 1.5% tween-80, 0.3% Vitamin C, and 20% propylene glycol (PG). Furthermore, the degradation rates of Z-ligustilide were found to conform to a rate equation following Weibull probability distribution within a range of degradation ratio, and the equation could be expressed as follow: ln ln (1/1-alpha) = ln k + m ln t. Where alpha is degradation ratio; t is time; m and k are constants relating to the degradation rate. The degradation rate will get greater as the increasing of parameter k. According to the degradation law obtained from the equation, the drug shelf-life (10% of active ingredient degraded, T90) in this vehicle was predicted to be more than 1.77 years at 25 degrees C through Arrehenius equation and accelerating experiments. The present investigation was undertaken to propose a kinetic treatment that may be applicable to any type of degradation of the active ingredient of pharmaceutical formulation, and also could provide a good foundation for the new drug development of Z-ligustilide, especially for injection formulation.


Binding of the specific ligand to Fc receptors on Trypanosoma cruzi increases the infective capacity of the parasite.
The infective capacity of Trypanosoma cruzi was significantly increased after treatment with monoclonal IgG1 antibodies, whether or not specific for the parasite; minimal or no change in infectivity was seen after treatment with IgG2a, IgG2b or IgG3 monoclonal antibodies. The stimulatory effect was evidenced by elevated numbers of trypanosomes invading mammalian host cells in vitro compared to parasites treated with medium alone. Greater infectivity was also induced by pure human Fc, suggesting a role for Fc receptors on the organism. This inference received support in the fact that protein A inhibited the stimulatory effect of Fc. In addition, Fc-treated parasites incubated with fluorescein-labelled F(ab')2 from goat anti-human IgG exhibited fluorescence detectable by both ultraviolet microscopy and flow cytometry. 125I-Fc binding to T. cruzi was found to be saturable at 0 degrees and was inhibited by cold Fc but not by bovine serum albumin (BSA) or orosomucoid. Interestingly, 125I-Fc binding was greater at 37 degrees and it was not saturable with the concentrations that did saturate at 0 degrees. Possibly, Fc might up-regulate expression of its own receptor and greater endocytosis could take place at 37 degrees. Significant increases in infectivity were detectable after a 40 min pretreatment with Fc--hinting that Fc could trigger a chain of biochemical events underlying the phenomenon--and were reversible, becoming undetectable 2 hr after Fc removal. The average number of Fc receptors per parasite, determined at 0 degrees (at which binding saturation was possible), was estimated as 5 x 10(5), the dissociation constant was of the order of 10(-6)-10(7)M. The present results define an important biological role for an Fc-binding T. cruzi surface component and expose the capacity of this organism to exploit even elements of the immune system in its quest to attain intracellular localization, required for multiplication.


Formation of soluble hexanuclear neptunium(IV) nanoclusters in aqueous solution: growth termination of actinide(IV) hydrous oxides by carboxylates.
Complexation of Np(IV) with several carboxylates (RCOO(-); R = H, CH(3), or CHR'NH(2); R' = H, CH(3), or CH(2)SH) in moderately acidic aqueous solutions was studied by using UV-vis-NIR and X-ray absorption spectroscopy. As the pH increased, all investigated carboxylates initiated formation of water-soluble hexanuclear complexes, Np(6)(μ-RCOO)(12)(μ(3)-O)(4)(μ(3)-OH)(4), in which the neighboring Np atoms are connected by RCOO(-)syn-syn bridges and the triangular faces of the Np(6) octahedron are capped with μ(3)-O(2-)/μ(3)-OH(-). The structure information of Np(6)(μ-RCOO)(12)(μ(3)-O)(4)(μ(3)-OH)(4) in aqueous solution was extracted from the extended X-ray absorption fine structure data: Np-O(2-) = 2.22-2.23 Å (coordination number N = 1.9-2.2), Np-O(RCOO(-)) and Np-OH(-) = 2.42-2.43 Å (N = 5.6-6.7 in total), Np···C(RCOO(-)) = 3.43 Å (N = 3.3-3.9), Np···Np(neighbor) = 3.80-3.82 Å (N = 3.6-4.0), and Np···Np(terminal) = 5.39-5.41 Å (N = 1.0-1.2). For the simpler carboxylates, the gross stability constants of Np(6)(μ-RCOO)(12)(μ(3)-O)(4)(μ(3)-OH)(4) and related monomers, Np(RCOO)(OH)(2)(+), were determined from the UV-vis-NIR titration data: when R = H, log β(6,12,-12) = 42.7 ± 1.2 and log β(1,1,-2) = 2.51 ± 0.05 at I = 0.62 M and 295 K; when R = CH(3), log β(6,12,-12) = 52.0 ± 0.7 and log β(1,1,-2) = 3.86 ± 0.03 at I = 0.66 M and 295 K.


Recall of three words after five minutes: its relationship to performance on neuropsychological memory tests.
This paper examines the statistical relationship between a memory screening test (recall of three words after 5 minutes, 3W5M) and an extensive battery of neuropsychological memory tests. Subjects were 142 consecutive referrals for neuropsychological examination and were given 3W5M in addition to measures of memory, language, and intelligence. Performance on 3W5M was categorized as either "impaired" or "unimpaired" in a manner consistent with judgement used in clinical settings. Linear discriminant function analysis was used to predict 3W5M performance from neuropsychological test scores. Results indicated that 3W5M was a relatively good screening measure, but that its sensitivity and utility as a measure of memory was significantly influenced by the nature of the neurological disorder. Significantly higher diagnostic accuracy rates were obtained in patient groups with clinically obvious memory impairment (e.g., dementia, pure amnesia) than in groups with less obvious memory impairment (e.g., closed head injury, psychiatric disorders). Implications for clinical memory testing, and for the relationship between screening and battery approaches to assessment, are discussed.


Classes of Individual Growth Trajectories of Religious Coping in Older Adulthood: Patterns and Predictors.
For many individuals, religion provides important cognitive resources for coping with stressors, especially in older adulthood. Although older adults are thought to make more use of these coping strategies than those at younger ages, less is known about how patterns of use change during the span of older adulthood. In a largely Christian sample of U.S. older adults, positive and negative religious coping were measured between 2 and 5 times over a period of 11 years (N = 1,075). Growth mixture modeling extracted latent classes of growth. The optimal solution for positive coping indicated a five-class structure (high, stable; high, declining moderately; high, declining rapidly; low, increasing; and low, stable) and the optimal negative coping solution had three classes (low, declining; low, increasing; and high, declining). Nominal logistic regression examined the relationship of individual characteristics with latent class. Education, religious commitment, religious attendance, and religious doubt were related to positive coping trajectory class. Only religious doubt was related to negative coping class.


Cost effectiveness of primary pegfilgrastim prophylaxis in patients with breast cancer at risk of febrile neutropenia.
Guidelines advise primary granulocyte colony-stimulating factor (G-CSF) prophylaxis during chemotherapy if risk of febrile neutropenia (FN) is more than 20%, but this comes with considerable costs. We investigated the incremental costs and effects between two treatment strategies of primary pegfilgrastim prophylaxis. Our economic evaluation used a health care perspective and was based on a randomized study in patients with breast cancer with increased risk of FN, comparing primary G-CSF prophylaxis throughout all chemotherapy cycles (G-CSF 1-6 cycles) with prophylaxis during the first two cycles only (G-CSF 1-2 cycles). Primary outcome was cost effectiveness expressed as costs per patient with episodes of FN prevented. The incidence of FN increased from 10% in the G-CSF 1 to 6 cycles study arm (eight of 84 patients) to 36% in the G-CSF 1 to 2 cycles study arm (30 of 83 patients), whereas the mean total costs decreased from € 20,658 (95% CI, € 20,049 to € 21,247) to € 17,168 (95% CI € 16,239 to € 18,029) per patient, respectively. Chemotherapy and G-CSF determined 80% of the total costs. As expected, FN-related costs were higher in the G-CSF 1 to 2 cycles arm. The incremental cost effectiveness ratio for the G-CSF 1 to 6 cycles arm compared with the G-CSF 1 to 2 cycles arm was € 13,112 per patient with episodes of FN prevented. We conclude that G-CSF prophylaxis throughout all chemotherapy cycles is more effective, but more costly, compared with prophylaxis limited to the first two cycles. Whether G-CSF prophylaxis throughout all chemotherapy cycles is considered cost effective depends on the willingness to pay per patient with episodes of FN prevented.


A Cell-Based Platform for the Investigation of Immunoproteasome Subunit β5i Expression and Biology of β5i-Containing Proteasomes.
The degradation of most intracellular proteins is a dynamic and tightly regulated process performed by proteasomes. To date, different forms of proteasomes have been identified. Currently the role of non-constitutive proteasomes (immunoproteasomes (iPs) and intermediate proteasomes (intPs)) has attracted special attention. Here, using a CRISPR-Cas9 nickase technology, four cell lines: histiocytic lymphoma, colorectal adenocarcinoma, cervix adenocarcinoma, and hepatocarcinoma were modified to express proteasomes with mCherry-tagged β5i subunit, which is a catalytic subunit of iPs and intPs. Importantly, the expression of the chimeric gene in modified cells is under the control of endogenous regulatory mechanisms and is increased following IFN-γ and/or TNF-α stimulation. Fluorescent proteasomes retain catalytic activity and are distributed within the nucleus and cytoplasm. RNAseq reveals marginal differences in gene expression profiles between the modified and wild-type cell lines. Predominant metabolic pathways and patterns of expressed receptors were identified for each cell line. Using established cell lines, we demonstrated that anti-cancer drugs Ruxolitinib, Vincristine and Gefitinib stimulated the expression of β5i-containing proteasomes, which might affect disease prognosis. Taken together, obtained cell lines can be used as a platform for real-time studies of immunoproteasome gene expression, localization of iPs and intPs, interaction of non-constitutive proteasomes with other proteins, proteasome trafficking and many other aspects of proteasome biology in living cells. Moreover, the established platform might be especially useful for fast and large-scale experiments intended to evaluate the effects of different conditions including treatment with various drugs and compounds on the proteasome pool.


Circulating miRNAs as Novel Diagnostic Biomarkers in Nonalcoholic Fatty Liver Disease: A Systematic Review and Meta-Analysis.
Recent studies have indicated that circulating miRNAs could serve as accurate biomarkers for diagnosing nonalcoholic fatty liver disease (NAFLD). We aimed to assess the evidence on the probability of circulating miRNAs as new diagnostic biomarkers in patients with NAFLD. We comprehensively retrieved relevant English literature from the databases of PubMed, Embase, and the Cochrane Library from 2000 to 1 January 2019. The diagnostic accuracy of circulating miRNAs as markers for NAFLD was analyzed. Moreover, we evaluated the methodological quality of the included article. STATA was applied to perform statistical analyses. In this meta-analysis, 17 studies that enrolled 1408 patients of NAFLD and 926 healthy people from 6 articles were analyzed. We constructed a summary receiver-operating characteristic (SROC) curve of all circulating miRNAs, and the area under the curve (AUC) was 0.83, with the pooled sensitivity (SEN) 0.70 and the pooled specificity (SPE) 0.82 in distinguishing patients with NAFLD from healthy controls. Among them, miR-122 showed high diagnostic accuracy, with the diagnostic index of pooled SEN, SPE, and AUC being 0.88, 0.66, and 0.86, respectively. We then performed subgroup analyses based on the mode of miRNA regulation, countries, miRNA profiling, sample size, and male proportion. We then did a regression analysis and found the cause of heterogeneity might be miRNA profiling. Finally, publication bias was not found, and Fagan's nomogram showed valuable clinical utility. Circulating miRNAs, especially miR-122, might be promising diagnostic biomarkers for NAFLD with high-accuracy, and more large-sample studies are required to support the above findings in the future.


Controlling burst and final drug release times from porous polylactide devices derived from co-continuous polymer blends.
This work has demonstrated that it is possible to exercise a wide range of control over both the initial burst release and the final drug release times from porous polylactide (PLA) devices derived from cocontinuous polymer blends. Two strategies were used: a layer-by-layer polyelectrolyte surface deposition approach on the porous PLA surface and the application of a partially closed-cell protocol. A PLA porous substrate with a pore size of 1.5 microm, derived from a blend of PLA and polystyrene (PS) via selective solvent extraction of the PS phase, was used as the drug delivery device. The surface area and pore dimensions were examined via BET nitrogen adsorption and image analysis. Porous PLA substrates with 0, 3, and 5 layers of polyelectrolytes and with open areas of 100, 12, and 2% were studied both separately and in combination. In vitro release tests were performed to study the release profile of bovine serum albumin (BSA) from the devices via UV spectrophotometry. It is shown that, while both are important, surface modification is more dominant in controlling the release rate than the partially closed cell approach. When a five layer surface modification of the PLA and a partially closed cell approach (2% open area) are combined, denoted as the L5C sample, the synergy is dramatic with a 5x reduction in the first two hour burst release amount and a total release time that is extended by 123x as compared to the 100% open cell, surface unmodified, reference sample. The L5C sample ultimately releases 89% of the total BSA loaded, demonstrating the high level of interconnectivity of the microchannels in the porous PLA. The mechanism of release in this system is clearly diffusion controlled with well-defined concentration gradients, as measured by X-ray photoelectron spectroscopy (XPS), observed in the direction of release. These results point toward a diffusion mechanism combined with a sorption/desorption interaction of the BSA with the modified PLA surface.


Depressive deficits in memory: implications for memory improvement following traumatic brain injury.
Experimental findings of memory impairment in depression are reviewed, and implications for rehabilitation are discussed. Impairments typically occur when the structure of the task is insufficient to constrain the nature of the procedures to be used. When such constraint is provided, or when the task is not particularly sensitive to strategic processing, depression-related impairments are rarely observed.


Tracheal reconstructions.
Surgical reconstruction of the trachea is a relatively complex procedure. We had 20 cases of tracheal stenosis. We have a modest experience of 16 tracheal reconstructions for acquired tracheal stenosis. Two patients underwent laser treatment while another two died before any intervention. The majority of these cases were a result of prolonged ventilation (14 cases), following organophosphorous poisoning (11 cases), Guillain-Barré syndrome, bullet injury, fat embolism and surprisingly only one tumor, a case of mucoepidermoid carcinoma, who had a very unusual presentation. There were 12 males and 4 females in this series, age ranging from 12-35 years. The duration of ventilation ranged from 1-21 days and the interval from decannulation to development of stridor was between 5-34 days. Six of them were approached by the cervical route, 5 by thoracotomy and cervical approach, 2 via median sternotomy and 3 by thoracotomy alone. Five of them required an additional laryngeal drop and 1 required pericardiotomy and release of pulmonary veins to gain additional length. The excised segments of trachea measured 3 to 5 cms in length. All were end to end anastomosis with interrupted Vicryl sutures. We have had no experience with stents or prosthetic tubes. Three patients developed anastomotic leaks which were controlled conservatively. Almost all of them required postoperative tracheo-bronchial suctioning with fibreoptic bronchoscope. We had one death in this series due to sepsis.


Rapid vasoregulatory mechanisms in exercising human skeletal muscle: dynamic response to repeated changes in contraction intensity.
We tested the hypothesis that vasoregulatory mechanisms exist in humans that can rapidly adjust muscle blood flow to repeated increases and decreases in exercise intensity. Six men and seven women (age, 24.4+/-1.3 yr) performed continuous dynamic forearm handgrip contractions (1- to 2-s contraction-to-relaxation duty cycle) during repeated step increases and decreases in contraction intensity. Three step change oscillation protocols were examined: Slow (7 contractions per contraction intensityx10 steps); Fast (2 contractions per contraction intensityx15 steps); and Very Fast (1 contraction per contraction intensityx15 steps). Forearm blood flow (FBF; Doppler and echo ultrasonography), heart rate (ECG), and mean arterial pressure (arterial tonometry) were examined for the equivalent of a cardiac cycle during each relaxation phase (FBFrelax). Mean arterial pressure and heart rate did not change during repeated step changes (P=0.352 and P=0.190). For both Slow and Fast conditions, relaxation phase FBFrelax adjusted immediately and repeatedly to both increases and decreases in contraction intensity, and the magnitude and time course of FBFrelax changes were virtually identical. For the Very Fast condition, FBFrelax increased with the first contraction and thereafter slowly increased over the course of repeated contraction intensity oscillations. We conclude that vasoregulatory mechanisms exist in human skeletal muscle that are capable of rapidly and repeatedly adjusting muscle blood flow with ongoing step changes in contraction intensity. Importantly, they demonstrate symmetry in response magnitude and time course with increasing versus decreasing contraction intensity but cannot adjust to very fast exercise intensity oscillations.


Adoption and maintenance of physical activity: Planning interventions in young, middle-aged, and older adults.
Young, middle-aged, and older adults in orthopaedic outpatient rehabilitation (N = 373) were randomly assigned to either an interviewer-assisted or a standard-care self-administered planning intervention. Physical activity planning consisted of specifying action plans to facilitate action initiation, and coping plans to overcome barriers. The interviewer-assisted condition led to more complete action plans and a longer duration of physical activities up to six months after discharge. Regarding coping planning, older and middle-aged adults benefited more from interviewer-assisted planning while younger adults benefited more from self-administered planning. Planning as such was found to be an effective tool for enactment irrespective of chronological age. The delayed effect of coping planning on enactment suggests that coping planning is important for long-term maintenance.


Hydraena (Hydraenopsis) ateneo, new species (Coleoptera, Hydraenidae) and other aquatic Polyphaga from a small habitat patch in a highly urbanized landscape of Metro Manila, Philippines.
Seven species of Hydraenidae, Hydrophilidae and Elmidae are recorded from temporary freshwater habitats at the Ateneo de Manila University Campus in the metropolitan area of Manila, Philippines. They were identified as Enochrus (Lumetus) fragiloides d'Orchymont, Helochares (Hydrobaticus) lepidus d'Orchymont, Helochares (Helochares) pallens (MacLeay), Hydraena (Hydraenopsis) scabra d'Orchymont, Hydraena (Hydraenopsis) palawanensis Freitag & Jäch (new record for Luzon Island), Stenelmis sp. A further hydraenid species was unknown to science and is newly described: Hydraena (Hydraenopsis) ateneo Freitag, sp. n. Aedeagus, gonocoxite, spermatheca, and female tergite X are illustrated by computer-based line drawings. Habitus images of all three Hydraena Kugelann species recorded and a checklist of the Philippine Hydraena are provided. The presence of these seven species in the Ateneo campus is briefly discussed in regard to the area's history. Measures to maintain and extend semi-natural islands of biodiversity in urban areas are suggested.


Lithium zincopyrophosphate, Li(2)Zn(3)(P(2)O(7))(2).
The title compound, dilithium(I) trizinc(II) bis[diphosphate(4-)], is the first quaternary lithium zincopyrophosphate in the Li-Zn-P-O system. It features zigzag chains running along c, which are built up from edge-sharing [ZnO(5)] trigonal bipyramids. One of the two independent Zn sites is fully occupied, whereas the other is statistically disordered by Zn(2+) and Li(+) cations, although the two Zn sites have similar coordination environments. Li(+) cations occupy a four-coordinated independent site with an occupancy factor of 0.5, as well as being disordered on the partially occupied five-coordinated Zn site with a Zn(2+)/Li(+) ratio of 1:1.


Stabilized beta-catenin immortalizes colonic epithelial cells.
The majority of colonic neoplasias contain mutations in either the adenomatous polyposis coli or the beta-catenin (beta-cat) gene, both of which result in elevated levels of cytoplasmic beta-cat. The oncogenic activity of beta-cat has been explored in vivo and in vitro with conflicting results. We tested the hypothesis that beta-cat is capable of immortalizing and transforming cultured epithelial cells that represent the precursors to colon cancer. A truncated form of beta-cat (deltaN89) was stably expressed in murine colonic epithelial cells that were conditionally immortalized by temperature-sensitive T antigen expression and contained a mutant ApcMin allele [Immorto-Min colonic epithelium (IMCE)]. IMCE cells, grown under nonpermissive conditions, were immortalized by expression of the truncated beta-cat protein as determined by sustained growth in culture and escape from senescence as measured by endogenous beta-galactosidase activity. IMCE neo cells at nonpermissive conditions underwent extensive apoptosis, an effect that was blocked by the expression of deltaN89 beta-catenin. IMCE beta-cat cells had significantly lower p19 and p53 protein levels compared to IMCE neo cells, suggesting that alterations in these two key genes may mediate the effects of beta-cat on both cellular senescence and apoptosis. IMCE beta-cat cells were also transformed as determined by growth in the absence of serum, anchorage-independent growth, and sustained tumor growth in nude mice. Stable beta-cat-expressing populations could not be generated in conditionally immortalized colonic epithelia cells with a wild-type Apc background. These studies demonstrated the immortalizing activity of stabilized beta-cat for the first time and extend the transforming ability of mutated beta-cat to a cell line representing a precursor to colorectal cancer.


[Distribution of human papillomavirus (HPV) among HPV positive cervical adenocarcinoma cases detected by laser capture microdissection(LCM)].
To investigate the distribution of human papillomavirus (HPV) in the diseased areas cut from HPV-positive cervical adenocarcinoma (ADC) detected by laser capture microdissection (LCM). Paraffin-embedded specimens diagnosed as ADC between 2005 and 2010 were collected from 9 hospitals in 7 regions across China. HPV genotyping was conducted on paraffin sections using sandwich technique and LCM in order to identify HPV infection in the tumor tissues. HE and p16 immunohistochemistry staining were performed to make histological diagnosis. A total of 169 cervical adenocarcinoma cases were recruited, including 94 cases of mucinous adenocarcinoma (ADC-CX), 9 cases of adenosquamous carcinoma (ASC), 19 cases of minimal deviation adenocarcinoma (ADC-MIN), 14 cases of clear cell adenocarcinoma (ADC-CC), 8 cases of endometrioid adenocarcinoma (ADC-ENDO), 9 cases of serous adenocarcinoma (ADC-SER) and 16 cases of adenocarcinoma not otherwise specified (ADC-NOS). Fourteen types of high risk HPV were detected in the whole tissue section (WTS). HPV16 was the most common type, and the second was HPV18 and HPV52, respectively. Compared with WTS, the HPV-positive rate detected by LCM was lower. The HPV positive rates were significantly different among different subtypes of cervical adenocarcinoma (P<0.001). After LCM, the HPV positive rate was 50.8% and 66.7% in the single infection and multiple, infection groups respectively (P=0.14). The positive rates of p16 was significantly different among different subtypes of cervical adenocarcinoma (P<0.001). p16-positive rate was 73.9% in the HPV-positive samples after LCM, significantly higher than the 38.5% of negative samples (P<0.001). Laser capture dissection technique can more precisely reflect the HPV distribution in cervical adenocarcinomas. The etiological association between HPV infection and cervical adenocarcinoma occurrence is not as close as that reported in the literature.


Splenectomy for people with thalassaemia major or intermedia.
Thalassaemia is a genetic disorder of the haemoglobin protein in red blood cells. It has been historically classified into thalassaemia minor, intermedia and major, depending on the genetic defect and severity of the disease. The clinical presentation of β-thalassaemia varies widely from a mild asymptomatic form in thalassaemia minor, to a severe disease in thalassaemia major where individuals are dependant on life-long blood transfusions. The hallmark of thalassaemia syndromes is the production of defective red blood cells that are removed by the spleen resulting in an enlarged hyperfunctioning spleen (splenomegaly). Removal of the spleen may thus prolong red blood cell survival by reducing the amount of red blood cells removed from circulation and may ultimately result in the reduced need for blood transfusions. To assess the efficacy and safety of splenectomy in people with β-thalassaemia major or intermedia. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Review Group's Haemoglobinopathies Trials Register, compiled from searches of electronic databases and the handsearching of journals and conference abstract books. We also searched online trial registries and the reference lists of relevant articles and reviews (27 July 2018).Date of the most recent search of the Group's trials register: 02 August 2019. We included randomised controlled and quasi-randomised controlled studies of people of any age with thalassaemia major or intermedia, evaluating splenectomy in comparison to conservative treatment (transfusion therapy and iron chelation) or other forms of splenectomy compared to each other (laparoscopic, open, radio-frequency). Two authors independently selected and extracted data from the single included study using a customised data extraction form and assessed the risk of bias. The quality of the evidence was assessed using GRADE. One study, including 28 participants was included in the review; the results were described, primarily, in a narrative manner. This study assessed the feasibility of splenectomy using a laparoscopic approach versus open surgery. Given the lack of detail regarding the study methods beyond randomisation, the overall risk of bias for this study was unclear. The study was carried out over a period of 3.5 years, with each participant followed up only until discharge (less than one week after the intervention); it did not assess the majority of the outcomes outlined in this review (including two of the three primary outcomes, frequency of transfusion and quality of life). A total of three serious post-operative adverse events (the review's third primary outcome) were reported in the laparoscopic splenectomy group (one case of atelectasis and two cases of bleeding), compared to two events of atelectasis in the open surgery group; however, there were no significant differences between the groups for either atelectasis, risk ratio (RR) 0.50 (95% confidence interval (CI) 0.05 to 4.90) or for bleeding, RR 5.00 (95% CI 0.26 to 95.61) (very low-quality evidence). In addition, the study also reported three serious cases of intra-operative bleeding in the laparoscopic group which mandated conversion to open surgery, although the difference between groups was not statistically significant, RR 7.00 (95% CI 0.39 to 124.14) (very low-quality evidence). These effect estimates are based on very small numbers and hence are unreliable and imprecise. From this small study, there appeared to be an advantage for the laparoscopic approach, in terms of post-operative hospital stay, although the group difference was not large (median difference of 1.5 days, P = 0.03). The review was unable to find good quality evidence, in the form of randomised controlled studies, regarding the efficacy of splenectomy for treating thalassaemia major or intermedia. The single included study provided little information about the efficacy of splenectomy, and compared open surgery and laparoscopic methods. Further studies need to evaluate the long-term effectiveness of splenectomy and the comparative advantages of surgical methods. Due to a lack of high quality evidence from randomised controlled studies, well-conducted observational studies may be used to answer this question.


Biologic effects of radiofrequency thermal ablation on non-small cell lung cancer: results of a pilot study.
Radiofrequency ablation of lung tumors is an emerging technique with promising results. To achieve more information on its effects on pulmonary parenchyma and primary tumors, and to assess its efficacy in determining complete necrosis of the lesion, we led a pilot study consisting of thermal ablation followed by surgical resection. Ten patients with early stage non-small cell lung cancer were enrolled in the study. In 5 patients radiofrequency ablation was achieved through thoracotomy just before the surgical resection. In another 5 patients radiofrequency ablation was performed percutaneously, under computed tomography guidance, and the surgical resection was performed after 15 days. Nine of the 10 patients enrolled in the study were available for analysis. There were 8 men and 1 woman with a mean age of 65.5 years. In all cases histologic diagnosis was available before radiofrequency ablation; adenocarcinoma was identified in 5 patients, and squamous cell carcinoma was identified in 4 patients. Microscopic examination showed complete necrosis of the tumor in 6 of 9 cases. No anatomopathologic alterations were seen in surrounding distant parenchyma. Our study confirms the safety of radiofrequency ablation, particularly underlining the possibility of performing a thermal lesion in the lung in a controlled manner. It also assesses that radiofrequency ablation may be effective in the local control of primary lung cancer. However, surgery still represents the mainstay of treatment of non-small cell lung cancer, and radiofrequency ablation must be reserved for high-risk patients.


Relations Between Bilingualism and Autistic-Like Traits in a General Population Sample of Primary School Children.
Some evidence suggests that bilingualism improves communication and cognitive skills which are often impaired in autism. However, diagnosing autism in bilinguals may suffer a cultural bias, which can affect the investigation of bilingualism and autism. Therefore, the current study investigates relations between autistic-like traits (ALTs) and bilingualism in a general population sample of 394 children (M age = 8;3). Within the high-scoring group on the ALT measure, monolinguals had significantly higher ALT scores than bilinguals. There were no differences between monolinguals and bilinguals in the low-scoring group. Across the whole sample, age and structural language skills accounted for 35% variance in ALTs, while bilingualism had no effect on ALTs. Furthermore, structural language skills explained more variance in ALTs among bilinguals than among monolinguals.


Identification of differentially expressed genes in hepatocellular carcinoma with cDNA microarrays.
Genes expressed in hepatocellular carcinoma (HCC) were analyzed using cDNA microarrays to clarify gene abnormalities in HCC. mRNA was extracted from cancerous and noncancerous tissues of 10 patients with HCC, cDNA labeled with Cy5 and Cy3 fluorescence was prepared, and it was hybridized for each patient with a cDNA microarray consisting of 1,080 elements (930 unique genes). The mRNA expression rate of each element in HCC was evaluated using the level of mRNA expression in noncancerous tissue in each patient as a reference. The expression of 10 genes was enhanced 2 times or more in HCC cancerous tissue compared with noncancerous tissue in 5 or more of the 10 patients. In contrast, 9 genes were expressed at half the level or less in HCC cancerous tissue compared with noncancerous tissue. When hierarchical clustering was performed to identify genes related to clinical phenotypes of the patients, 22 genes showed changes associated with the degree of differentiation of HCC. Thirteen of these genes were transcriptional factors or tissue-specific expression proteins related to cell differentiation or development. Our present analysis clarified a number of genes that characterize HCC. This information based on examination of clinical samples is considered to be useful for clarification of the mechanism of hepatocarcinogenesis and the diagnosis and treatment of HCC.


Improved Lycopene Production from Different Substrates by Mated Fermentation of Blakeslea Trispora.
The production of lycopene from different substrates by Blakeslea trispora in fermentation was investigated. Lycopene productions from 4 and 6% glucose (pH 6.5) in shake flask fermentation were 77.7 and 28.1 mg L-1. Increasing the glucose concentration to 6% resulted in a decrease in lycopene production by 36.2%. A maximum lycopene concentration of 944.8 mg L-1 was detected with 4% glucose supplemented with 1.0 % sunflower oil in fermentor studies. Lycopene productions in the presence of sunflower and corn oils in the fermentor were 12.2 and 11.1 times higher, respectively, then without oil from 4 % glucose in a shake flask. Lycopene production from orange peel was two times higher in the fermentor than in the shake flask. Zygospores of B. trispora are the morphological forms, which are responsible for the production of the lycopene. The highest level of zygospores was correlated with the highest amount of intracellular lycopene in the total biomass dry weight. The media containing only orange powder (1%) gave a 4.9 mg L-1 lycopene production in a fermentor. The biosynthesis of lycopene has been started in most cases simultaneously in the early growth phase even in trace amounts. Maximum lycopene concentration was obtained when the medium was supplied with sunflower and corn oils. There is an indirect relationship between biomass and lycopene concentration.


Complete characterisation of thermally treated sludges.
As disposal options for sludges become more difficult to find and more expensive to operate, those relying on some form of thermal treatment are becoming more commercially attractive. The incentive, especially for more heavily populated countries, is the production of treated solids suitable for beneficial application to land. It is the level of treatment required to achieve this cost effectively which has been the focus of much of the research in this field. There are now several commercially available plants designed to thermally treat sludges and many have been in full scale operation for a number of years. One of the auxiliary claims often made by proponents of such treatments, is that the resultant sludges are more amenable to dewatering and therefore easier and cheaper to handle. In this work we have used a novel filtration rig to obtain complete sets of filtration data for different sludges. Historically sludge characterisation has been achieved by measurement of an empirical Capillary Suction Time (CST) parameter, however with the new filtration apparatus it is now possible to obtain fundamental sludge characteristics across a wide range of volume fractions in hours not days. We have applied this technology to characterise different sludge samples before and after thermal treatment under different sets of operating conditions (pH, temperature and pressure) to simulate some of the commercially available thermal treatment technologies. We have also examined the effects of various chemical oxidants (hydrogen peroxide and Fenton's reagent) used as pretreatments to the thermal process and attempted to compare their cost efficiencies. The results show that the physical structure of the sludge is irreversibly altered by decreasing the pH or heating to temperatures in excess of 150 degrees C in a way which significantly enhances the dewaterability of the material. The treated sludge not only has a higher permeability but also has a higher final per cent solids (often in the autothermal region). In contrast the amounts of chemical additives used in pretreatment to achieve similar effects are cost prohibitive.


Development of an Indirect ELISA to Detect African Swine Fever Virus pp62 Protein-Specific Antibodies.
African swine fever (ASF) is a highly detrimental viral disease caused by African swine fever virus (ASFV). The occurrence and prevalence of this disease have become a serious threat to the global swine industry and national economies. At present, the detection volume of African swine fever is huge, more sensitive and accurate detection techniques are needed for the market. pp62 protein, as a protein in the late stage of infection, has strong antigenicity and a high corresponding antibody titer in infected pigs. In this study, the CP530R gene was cloned into expression vector pET-28a to construct a prokaryotic expression plasmid, which was induced by IPTG to express soluble pp62 protein. Western blot analysis showed that it had great reactivity. Using the purified recombinant protein as an antigen, an indirect ELISA method for detecting ASFV antibody was established. The method was specific only to ASFV-positive serum, 1:1600 diluted positive serum could still be detected, and the coefficients of variation (CV) of the intra assay and inter assay were both <10%. It turns out that the assays had excellent specificity, sensitivity, and repeatability. This provides an accurate, rapid, and economical method for the detection of ASFV antibody in clinical pig serum samples.


Mechanism of augmented duodenal HCO(3)(-) secretion after elevation of luminal CO(2).
The proximal duodenum is exposed to extreme elevations of P(CO(2)) because of the continuous mixture of secreted HCO(3)(-) with gastric acid. These elevations (up to 80 kPa) are likely to place the mucosal cells under severe acid stress. Furthermore, we hypothesized that, unlike most other cells, the principal source of CO(2) for duodenal epithelial cells is from the lumen. We hence examined the effect of elevated luminal P(CO(2)) on duodenal HCO(3)(-) secretion (DBS) in the rat. DBS was measured by the pH-stat method. For CO(2) challenge, the duodenum was superfused with a high Pco(2) solution. Intracellular pH (pH(i)) of duodenal epithelial cells was measured by ratio microfluorometry. CO(2) challenge, but not isohydric solutions, strongly increased DBS to approximately two times basal for up to 1 h. Preperfusion of the membrane-permeant carbonic anhydrase inhibitor methazolamide, or continuous exposure with indomethacin, fully inhibited CO(2)-augmented DBS. Dimethyl amiloride (0.1 mM), an inhibitor of the basolateral sodium-hydrogen exchanger 1, also inhibited CO(2)-augumented DBS, although S-3226, a specific inhibitor of apical sodium-hydrogen exchanger 3, did not. DIDS, an inhibitor of basolateral sodium-HCO(3)(-) cotransporter, also inhibited CO(2)-augemented DBS, as did the anion channel inhibitor 5-nitro-2-(3-phenylpropylamino) benzoic acid. CO(2) decreased epithelial cell pH(i), followed by an overshoot after removal of the CO(2) solution. We conclude that luminal CO(2) diffused in the duodenal epithelial cells and was converted to H(+) and HCO(3)(-) by carbonic anhydrase. H(+) initially exited the cell, followed by secretion of HCO(3)(-). Secretion was dependent on a functioning basolateral sodium/proton exchanger, a functioning basolateral HCO(3)(-) uptake mechanism, and submucosal prostaglandin generation and facilitated hydration of CO(2) into HCO(3)(-) and H(+).


Potential of Polyphenols to Restore SIRT1 and NAD+ Metabolism in Renal Disease.
SIRT1 is an NAD+-dependent class III histone deacetylase that is abundantly expressed in the kidney, where it modulates gene expression, apoptosis, energy homeostasis, autophagy, acute stress responses, and mitochondrial biogenesis. Alterations in SIRT1 activity and NAD+ metabolism are frequently observed in acute and chronic kidney diseases of diverse origins, including obesity and diabetes. Nevertheless, in vitro and in vivo studies and clinical trials with humans show that the SIRT1-activating compounds derived from natural sources, such as polyphenols found in fruits, vegetables, and plants, including resveratrol, quercetin, and isoflavones, can prevent disease and be part of treatments for a wide variety of diseases. Here, we summarize the roles of SIRT1 and NAD+ metabolism in renal pathophysiology and provide an overview of polyphenols that have the potential to restore SIRT1 and NAD+ metabolism in renal diseases.


Stability of ondansetron hydrochloride injection in extemporaneously prepared oral solutions.
The stability of ondansetron hydrochloride in extemporaneously prepared oral solutions containing orange juice, cola, or cherry syrup was determined. Solutions were prepared by adding ondansetron hydrochloride to orange juice, cola, or cherry syrup to produce ondansetron concentrations of 0.267 and 0.067 mg/mL in orange juice or cola and 0.533 mg/mL in cherry syrup. The ondansetron concentration in orange juice and cola solutions was assayed at the time of preparation and at 30 minutes and one hour. The cherry syrup solution was stored at both 3-5 and 25-27 degrees C, with the ondansetron concentration being determined at the time of preparation and daily for seven days. All the solutions were prepared in triplicate. Ondansetron concentrations were measured by stability-indicating high-performance liquid chromatography. At each time interval, the mean ondansetron concentration remained > or = 97% of the initial measurement for all solutions. The appearance and color of the solutions did not change. Ondansetron hydrochloride was stable for at least one hour in orange juice or cola and at least seven days in cherry syrup.


A QTL on the short arm of wheat (Triticum aestivum L.) chromosome 3B affects the stability of grain weight in plants exposed to a brief heat shock early in grain filling.
Molecular markers and knowledge of traits associated with heat tolerance are likely to provide breeders with a more efficient means of selecting wheat varieties able to maintain grain size after heat waves during early grain filling. A population of 144 doubled haploids derived from a cross between the Australian wheat varieties Drysdale and Waagan was mapped using the wheat Illumina iSelect 9,000 feature single nucleotide polymorphism marker array and used to detect quantitative trait loci for heat tolerance of final single grain weight and related traits. Plants were subjected to a 3 d heat treatment (37 °C/27 °C day/night) in a growth chamber at 10 d after anthesis and trait responses calculated by comparison to untreated control plants. A locus for single grain weight stability was detected on the short arm of chromosome 3B in both winter- and autumn-sown experiments, determining up to 2.5 mg difference in heat-induced single grain weight loss. In one of the experiments, a locus with a weaker effect on grain weight stability was detected on chromosome 6B. Among the traits measured, the rate of flag leaf chlorophyll loss over the course of the heat treatment and reduction in shoot weight due to heat were indicators of loci with significant grain weight tolerance effects, with alleles for grain weight stability also conferring stability of chlorophyll ('stay-green') and shoot weight. Chlorophyll loss during the treatment, requiring only two non-destructive readings to be taken, directly before and after a heat event, may prove convenient for identifying heat tolerant germplasm. These results were consistent with grain filling being limited by assimilate supply from the heat-damaged photosynthetic apparatus, or alternatively, accelerated maturation in the grains that was correlated with leaf senescence responses merely due to common genetic control of senescence responses in the two organs. There was no evidence for a role of mobilized stem reserves (water soluble carbohydrates) in determining grain weight responses. Molecular markers for the 3B or 6B loci, or the facile measurement of chlorophyll loss over the heat treatment, could be used to assist identification of heat tolerant genotypes for breeding.


Rivaroxaban Once-Daily vs. Dose-Adjusted Vitamin K Antagonist on Biomarkers in Acute Decompensated Heart Failure and Atrial Fibrillation (ROAD HF-AF): Rationale and Design of an Investigator-Initiated Multicenter Randomized Prospective Open-Labeled Pilot Clinical Study.
Background: Clinical trials of non-vitamin K antagonist oral anticoagulants (NOACs) in patients with chronic heart failure and atrial fibrillation (AF) have demonstrated reduced risks of stroke and bleeding compared with vitamin K antagonists (VKAs). Here, we aim to assess the clinical efficacy and safety of rivaroxaban, a NOAC, compared with warfarin, a VKA, and the effects of rivaroxaban on cardiovascular biomarkers in patients with acute decompensated heart failure (ADHF) with reduced ejection fraction (≤40%) and AF. Methods: Rivaroxaban Once-daily vs. dose-adjusted vitamin K antagonist on biomarkers in Acute Decompensated Heart Failure and Atrial Fibrillation (ROAD HF-AF) is a randomized, open-labeled, controlled, prospective, multicenter pilot study designed to assess cardiovascular biomarkers and the safety of rivaroxaban (20 or 15 mg in patients with creatinine clearance 30-49 mL/min per day) compared with VKA (target international normalized range: 2-3) in 150 patients hospitalized with ADHF and AF. The primary endpoint is the change in circulating high-sensitivity cardiac troponin (hsTn) during hospitalization. The secondary endpoints are bleeding, hospital stay duration, in-hospital mortality, and changes in cardiovascular, renal, and thrombosis biomarkers. Patients will be followed for 180 days. Conclusion: We hypothesize that rivaroxaban will reduce myocardial injury and hemodynamic stress, as reflected by the biomarker status, within 72 h in patients with ADHF and AF, compared with VKA. We hope to facilitate future biomarker-based, large-scale outcome trials using NOACs in patients with ADHF and AF, based on the results of this multicenter, randomized, controlled study.


Chronologic change in the growth plate after radiofrequency-induced thermal injury: MRI-histologic correlation.
The purposes of this study were to assess chronologic changes in normal growth plate after radiofrequency-induced thermal injury and to evaluate the feasibility of MRI for revealing alteration of the growth plate. Radiofrequency ablation was performed on the right proximal tibia of 13 8-week-old New Zealand White rabbits. An 18-gauge cooled-tip electrode with a 5-mm active tip was placed distal to the physis under fluoroscopic guidance. MRI, including T1- and T2-weighted images, gradient-recalled echo images, and contrast-enhanced T1-weighted images, was performed 2, 4, and 12 weeks after ablation. Rabbits were sacrificed 2 (n = 4), 4 (n = 4), and 12 weeks (n = 5) after ablation. The sequential changes in the ablated zone, the injured physis, and the nonablated portion of the physis were correlated between MRI features and histologic results. Diameter of the nonenhancing lesion on MR images strongly correlated with the size of the region of coagulation necrosis at gross examination. The intraclass correlation coefficients were 0.98 and 0.94 for the long and short axes (p < 0.001). On gradient-recalled echo images, physial conspicuity was less in the injured physis than in the nonablated portion and less in the ablated than the control tibia. Physial conspicuity was graded for comparison with physial thickness at microscopic examination. The thickness of the physis was less in the ablated than in the control tibia 4 and 12 weeks after treatment (p < 0.05, paired Student t test). The cartilage column of the injured physis was delaminated 2 weeks after treatment, and a bone bridge through the injured physis was detected at 4 weeks. Radiofrequency-induced thermal injury causes early closure of the physis. MRI can depict the extent of radiofrequency-induced thermal injury and alterations in the physis that lead to early closure.


The Saccharomyces cerevisiae Dna2 can function as a sole nuclease in the processing of Okazaki fragments in DNA replication.
During DNA replication, synthesis of the lagging strand occurs in stretches termed Okazaki fragments. Before adjacent fragments are ligated, any flaps resulting from the displacement of the 5' DNA end of the Okazaki fragment must be cleaved. Previously, Dna2 was implicated to function upstream of flap endonuclease 1 (Fen1 or Rad27) in the processing of long flaps bound by the replication protein A (RPA). Here we show that Dna2 efficiently cleaves long DNA flaps exactly at or directly adjacent to the base. A fraction of the flaps cleaved by Dna2 can be immediately ligated. When coupled with DNA replication, the flap processing activity of Dna2 leads to a nearly complete Okazaki fragment maturation at sub-nanomolar Dna2 concentrations. Our results indicate that a subsequent nucleolytic activity of Fen1 is not required in most cases. In contrast Dna2 is completely incapable to cleave short flaps. We show that also Dna2, like Fen1, interacts with proliferating cell nuclear antigen (PCNA). We propose a model where Dna2 alone is responsible for cleaving of RPA-bound long flaps, while Fen1 or exonuclease 1 (Exo1) cleave short flaps. Our results argue that Dna2 can function in a separate, rather than in a Fen1-dependent pathway.


Randomized study of single-dose, three-day, and seven-day treatment of cystitis in women.
We evaluated the following five treatment regimens for acute cystitis in nonpregnant women: cefadroxil, 1,000 mg single-dose; cefadroxil, 500 mg twice a day for three days; cefadroxil, 500 mg twice a day for seven days; trimethoprim-sulfamethoxazole (TMP-SMZ), 320-1,600 mg single-dose, and TMP-SMZ, 160-800 mg twice a day for three days. At four weeks after the end of treatment, 25%, 58%, 70%, 65%, and 88% of patients, respectively, remained cured of infection. The results indicated that three-day treatment (1) might improve cure rates (over single-dose), (2) would reduce incidence of relapse (vs. single-dose), and (3) may be as curative as seven-day treatment. The results of the antibody-coated bacteria test did not predict treatment failure or relapse.


MMPI changes associated with therapeutic intervention: a migraine control study.
The investigation of personality traits of migraineurs with the Minnesota Multiphasic Personality Inventory (MMPI) is important research, but so far has led to diverse conclusions. This study aimed to investigate the influences of treatment intervention on the personality of migraineurs. Twenty-three Chinese patients (5 men, 18 women) with migraine (2 with aura, 21 without aura) were given the Chinese edition of the MMPI, before and after treatment, and were compared with 30 nonheadache healthy control subjects (6 men, 24 women). Statistical analyses were made among the three groups. The results revealed that patients in the pretreatment group with migraine had significantly higher scores on subtests of neuroticism (hypochondriasis, depression, hysteria) and schizophrenia. After treatment, the scores on subtests of hysteria, psychasthenia, and schizophrenia were remarkably lower (P < .05); the MMPI profile of the posttreatment group was within the reference range, but the scores of the neurotic scales were still higher than those of the healthy control group (P < .01). These results suggest that after treatment, disturbances in thinking, sentiment, and behavior were eliminated, and anxiety symptoms remarkably reduced, but some "migraine personality" characteristics remained and could influence the long-term results of treatment to some extent. It is suggested that management of migraine should include psychological intervention.


[Taxotere phase III trial on the first-line treatment of metastatic breast cancer]
OBJECTIVES: Doxorubicin and taxanes are the most effective agents in the treatment of advanced breast cancer. The aim of the study was to compare the efficacy of Doxorubicin (A) + Docetaxel (T) (AT) and standard Doxorubicin (A) + Cyclophosphamide (C) (AC) chemotherapy. MATERIALS AND METHODS: Results of first-line AT (50/75 mg/m2) and AC (60/600 mg/m2) D 1 q 3 wk, maximum of 8 cycles, were compared. Three Hungarian centers - Petz Aladár County Teaching Hospital, Gyôr, St.Margit Hospital, Budapest, and BAZ County Hospital, Miskolc, with 33 patients participated the international, phase III randomized TAX 306 trial. Between June, 1996 and March, 1998, 429 metastatic breast cancer patients were enrolled in the study. Eligible patients were who had not received prior chemotherapy for advanced disease, and were anthracycline-naive. Objective response rate observed in the AT arm was significantly higher than in the AC arm (ORR: 60% vs. 47%, p=0.008). Time to progression was longer in the AT group (37.1 weeks vs. 31.9 weeks, p=0.0153). Except for higher incidence of neutropenia not requiring dose modification in the AT arm, there were no major differences concerning toxicity. T did not enhance cardiac toxicity induced by A. CONCLUSION: AT results in significantly higher response rate and longer time to progression than AC in advanced breast cancer, even in patients with unfavourable prognosis.


Deafness, culture, and choice.
The recent controversy surrounding the choice, by a deaf lesbian couple, to have children who were themselves deaf, has focused attention on the ethics of choosing (apparent) disabilities for children. Deaf activists argue that deafness is not a disability, but instead the constitutive condition of access to a rich culture. Being deaf carries disadvantages with it, but these are a product of discrimination, not of the condition itself. It is, however, implausible to think that all the disadvantages which stem from deafness are social in origin. Moreover, though it may be true that being deaf carries with it the important compensation of access to a rich culture, no physical condition is required for such access. Cultures are simply the kind of things to which we are born, and therefore to which the children of deaf parents, hearing or deaf, normally belong. Thus these parents are making a mistake in choosing deafness for their children. Given their own experience of isolation as children, however, it is a mistake which is understandable, and our reaction to them ought to be compassion, not condemnation.


Primary sequence and developmental expression pattern of mRNAs and protein for an alpha1 subunit of the sodium pump cloned from the neural plate of Xenopus laevis.
Expression of a catalytic alpha subunit of the sodium pump was followed in early Xenopus embryos for correlation with physiological experiments showing that the sodium pump controls cavity expansion and the differentiation of neurones from the neural plate. Two cDNAs (one full length, one partial) for alpha1 subunit isoforms were cloned from a neural plate stage Xenopus library and sequenced. Other isoforms were not detected. Temporal and spatial expression patterns for alpha1 subunit transcripts and protein revealed extensive developmental regulation. At all stages, cells involved in cavity generation (outer ectoderm and cells lining the archenteron) expressed alpha1, transcripts with protein confined to the lateral and basal membranes. Before gastrulation, transcript levels were low and predominantly in animal cells. During gastrulation, alpha1 mRNAs rose significantly. Transcripts and protein were down-regulated in future outer neural plate cells as the mesoderm invaginated. Protein appeared at the blastopore on apical surfaces of lip cells and apposing surfaces of invaginating cells, suggesting that the Na pump opposes entry of fluid. In early neurulae, alpha1 mRNAs rose sharply. Transcript expression remained low in outer neural plate cells and increased in the endoderm, and protein appeared in the notochord. In midneurulae, transcripts returned in outer neural plate cells. Protein expression appeared on basal surfaces of deep neural plate cells and the floor plate, matching physiological observations. After neural tube closure, transcripts were detected in all dorsal structures. Protein was retained in the notochord and floor plate, was eliminated from the outer layer of the neural tube, and appeared on ependymal cells. The results are discussed in relation to previous physiological observations.


Circulating Cell-Free mtDNA Contributes to AIM2 Inflammasome-Mediated Chronic Inflammation in Patients with Type 2 Diabetes.
Mitochondrial dysfunction has been implicated in the pathogenesis of insulin resistance and type 2 diabetes. Damaged mitochondria DNA (mtDNA) may have a role in regulating hyperglycemia during type 2 diabetes. Circulating cell-free mitochondria DNA (ccf-mtDNA) was found in serum and plasma from patients and has been linked to the prognosis factors in various human diseases. However, the role of ccf-mtDNA in chronic inflammation in type 2 diabetes is unclear. In this study, we hypothesized that the ccf-mtDNA levels are associated with chronic inflammation in patients with type 2 diabetes. The mtDNA levels were elevated in the plasma from patients with type 2 diabetes compared to healthy subjects. The elevated mtDNA levels were associated with interleukin-1 (IL-1)β levels in patients with type 2 diabetes. The mtDNA, from patients with type 2 diabetes, induced absent in melanoma 2 (AIM2) inflammasome-dependent caspase-1 activation and IL-1β and IL-18 secretion in macrophages. Our results suggest that the ccf-mtDNA might contribute to AIM2 inflammasome-mediated chronic inflammation in type 2 diabetes.


Association between the Three Polymorphisms of the Glucocorticoid Receptor Gene and the Early Clinical Outcome in Kidney Transplantation Patients.
Glucocorticoids are pivotal components of immunosuppressive regimens in solid organ transplantations. This study aimed to assess the possible association between the ER22/23EK, N363S, and Bcl1 polymorphisms, and short-term clinical outcomes, including acute rejection and delayed graft function (DGF), in kidney transplantation recipients. A case-control study was conducted in a two-year period on adults with transplanted kidneys, comprised of subjects without rejection (n=50, control) and those with documented rejection within one year after transplantation (n=50, case), between April 2017 and September 2018, in Shiraz, Iran. Demographic characteristics and clinical and paraclinical findings were gathered. The genotyping of the ER22/23EK, N363S, and Bcl1 polymorphisms was carried out via polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The association between the genotypes and DGF as well as rejection types was evaluated using either the Chi square test or Fisher exact test. A stepwise logistic regression analysis was conducted to determine the independent factors of acute rejection within the first year after transplantation. The study population consisted of 64 men and 36 women. The frequency of mutated alleles was 0.32 for G (Bcl1), 0.02 for S (N363S), and 0.065 for A (ER22/23EK). There was no significant association either between the studied polymorphisms and acute rejection or between the Bcl1 (P=0.17), N363S (P=0.99), and ER22/23EK (P=0.99) genotypes and DGF. The length of hospital stay after kidney transplantation was slightly more in N363N and ER22/23EK wild allele carriers. However, this difference was not statistically significant. Our data suggested no statistically significant association between the genotypes of the studied polymorphisms and early clinical outcomes after kidney transplantation.


Differentiation molecules of the equine trophoblast.
Monoclonal antibodies raised against horse placenta were tested using an indirect immunoperoxidase-labelling technique for reactivity with a panel of tissues from adult horses and conceptuses of various gestational ages. The pattern of reactivity of 4 of the antibodies (F67.1, F71.3, F71.7, F71.14) on trophoblastic tissues described unique antigenic phenotypes for the non-invasive trophoblast of the allantochorion, the invasive trophoblast of the chorionic girdle, and the mature endometrial cup cells, which are derived from the chorionic girdle. Two of the monoclonal antibodies (F67.1 and F71.3) reacted only with chorionic girdle and the endometrial cups. Antibody F71.7 labelled strongly the non-invasive allantochorion from Day 29 of gestation to term. However, F71.7 failed to label mature endometrial cups and stained chorionic girdle only weakly, suggesting that the ability of the girdle cells to synthesize the molecule identified by F71.7 was gradually lost after development of the girdle. Antibody F71.14 reacted with trophoblastic tissues from all stages of gestation tested, with the exception of chorionic girdle. The other 3 anti-trophoblast monoclonal antibodies (F71.1, F71.2 and F71.8) labelled trophoblast-derived tissues from all stages tested. When the monoclonal antibodies were tested on cultured fetal and placental cells from Day 33 conceptuses recovered non-surgically from pregnant mares, the reactivities of the monoclonal antibodies on cultured cells were mostly identical to their reactivities in situ on tissue samples of similar gestational age; F67.1 and F71.3 were strong, specific markers for chorionic girdle cells, and F71.7 labelled allantochorion weakly in vitro, but failed to label chorionic girdle cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Experimental observations of fractal landscape dynamics in a dense emulsion.
Many soft and biological materials display so-called 'soft glassy' dynamics; their constituents undergo anomalous random motions and complex cooperative rearrangements. A recent simulation model of one soft glassy material, a coarsening foam, suggested that the random motions of its bubbles are due to the system configuration moving over a fractal energy landscape in high-dimensional space. Here we show that the salient geometrical features of such high-dimensional fractal landscapes can be explored and reliably quantified, using empirical trajectory data from many degrees of freedom, in a model-free manner. For a mayonnaise-like dense emulsion, analysis of the observed trajectories of oil droplets quantitatively reproduces the high-dimensional fractal geometry of the configuration path and its associated local energy minima generated using a computational model. That geometry in turn drives the droplets' complex random motion observed in real space. Our results indicate that experimental studies can elucidate whether the similar dynamics in different soft and biological materials may also be due to fractal landscape dynamics.


An aberrant venous channel mimicking the perilabyrinthine cells in the petrous bone of a patient with vestibular schwannoma: illustrative case.
Anatomical variations, such as high jugular bulbs and air cell development in the petrosal bone, should be evaluated before surgery. Most bone defects in the internal auditory canal (IAC) posterior wall are observed in the perilabyrinthine cells. An aberrant vascular structure passing through the petrous bone is rare. A 48-year-old man presented with a right ear hearing disturbance. Magnetic resonance imaging revealed a 23-mm contrast-enhancing mass in the right cerebellopontine angle extending into the IAC, consistent with a right vestibular schwannoma. Preoperative bone window computed tomographic scans showed bone defects in the IAC posterior wall, which ran farther posteroinferiorly in the petrous bone, reaching the medial part of the jugular bulb. The tumor was accessed via a lateral suboccipital approach. There was no other major vein in the cerebellomedullary cistern, except for the vein running from the brain stem to the IAC posterior wall. To avoid complications due to venous congestion, the authors did not drill out the IAC posterior wall or remove the tumor in the IAC. Several aberrant veins in the petrous bone are primitive head sinus remnants. Although rare, their surgical implication is critical in patients with vestibular schwannomas.


Regulation of Fibroblast Cell Polarity by Src Tyrosine Kinase.
Src protein tyrosine kinases (SFKs) are a family of nonreceptor tyrosine kinases that are localized beneath the plasma membrane and are activated during cell adhesion, migration, and elongation. Due to their involvement in the activation of signal transduction cascades, SFKs have been suggested to play important roles in the determination of cell polarity during cell extension and elongation. However, the mechanism underlying Src-mediated polarity formation remains unclear. The present study was performed to investigate the mechanisms underlying Src-induced cell polarity formation and cell elongation using Src knockout fibroblasts (SYFs) together with an inhibitor of Src. Normal and Src knockout fibroblasts were also transfected with a wild-type c-Src, dominant negative c-Src, or constitutively active c-Src gene to analyze the changes in cell morphology. SYF cells cultured on a glass substrate elongated symmetrically into spindle-shaped cells, with the formation of focal adhesions at both ends of the cells. When normal fibroblasts were treated with Src Inhibitor No. 5, a selective inhibitor of Src tyrosine kinases, they elongated into symmetrical spindle-shaped cells, similar to SYF cells. These results suggest that cell polarity during extension and elongation may be regulated by SFKs and that the expression and regulation of Src are important for the formation of polarity during cell elongation.


[Bioelectromagnetic studies: an analysis of errors in evaluation of electromagnetic field exposure].
This paper presents a review of exposure systems used in biomedical studies. Typical exposure systems of the whole frequency range are described. The attention was paid to technical limitations of applicability of the discussed exposure systems and the problem of correlation between the study results obtained in closed (TEM cell) and free spaces. Computer simulations of power absorbed by investigated objects with different numerical methods were made. As presented estimations show, electromagnetic energy absorption may differ between individual objects. The difference is a function of the system dimension and the number of exposed objects. Moreover, the energy distribution in a single object is a function of the frequency, system dimension, and placement of the object in relation to field vectors, not mentioning individual properties of the object. The presented results undermine the interpretation of the results of biomedical investigations carried out with use of closed exposure systems. Even if all inaccuracies and approximations in presented estimations are taken into account, the significance of mutual interactions between tested objects and exposure systems raises no doubts. Therefore, an attempt should be made to re-interpret the results of biomedical studies and to develop a closer cooperation between physicians and engineers.


Knowledge and Oral Health Attitudes among Parents of Children with Congenital Heart Disease.
Congenital heart disease (CHD) is a devastating complex of diseases resulting from defects of development. It affects more than 1 of every 100 live births. Early preventive dental care should be adjusted to the special needs of these children in their first years of life. Knowledge of parental attitudes and experiences of dental care are therefore important. Aim : This study was done to assess the knowledge and attitudes among parents of children with congenital heart disease towards oral health and dental care. Materials and methods : Parents (n = 105) of children with congenital heart disease of an age ranging from 0 to 16 years were included in the study. A questionnaire was used to assess the knowledge and oral health attitudes. Results : The parents' knowledge was fair but the oral health attitudes were not very satisfactory. The parents in this study also recognized the importance of oral health for the well-being of rest of the body. Conclusion : The results of this study indicate that parents' and children's attitudes toward oral health and dental care need to be improved.


The cytokine responsive vascular smooth muscle cell enhancer of inducible nitric oxide synthase. Activation by nuclear factor-kappa B.
The production of inducible nitric oxide synthase (iNOS) within vascular smooth muscle (VSM) cells following exposure to proinflammatory cytokines is a major cause of the vasorelaxation and hypotension of septic shock. We have defined the cytokine-responsive element of the murine iNOS promoter, transfected into a VSM cell line, and the role of the NF-kappa B/Rel family of proteins in iNOS gene activation in these cells. The combination of interleukin-1, interferon-gamma, and tumor necrosis factor-alpha stimulates promoter activity by a factor of 8.1-fold; single cytokines show little activity, while pairs of cytokines produce an intermediate effect. Using a series of promoter deletion mutants, we have defined the cytokine-responsive element from position -890 to -1002; this region contains an NF-kappa B-binding site as well as a number of interferon response elements. Nuclear proteins from cytokine-stimulated VSM cells which bind to an oligonucleotide containing this kappa B site are composed of p65 together with an unidentified protein of 50 kDa, which is not a known Rel family member. A promoter mutant with a 2-base pair change within this kappa B site, which abolishes NF-kappa B binding, has an activity of only approximately 34% (S.E. +/- 1.5) of the wild-type promoter. In addition, protein binding to this site is abolished by a specific inhibitor of NF-kappa B activation, which also abrogates iNOS activity. Residual inducibility in such mutant promoters is attributable to the presence of an independently functioning downstream kappa B site (-85 to -75). The mechanism by which NF-kappa B activates the iNOS promoter in VSM cells in response to cytokines appears to be markedly different to that operative in macrophages in response to lipopolysaccharide.


Evidence for keratin proteins in normal and abnormal human meibomian fluids.
Hyperkeratinization of meibomian glands has been postulated to cause gland dysfunction. Recent investigations on rabbits show that keratin proteins are indeed present in the meibomian fluids of these animals. In this report we present our findings on the presence of these water-insoluble proteins in human meibomian secretions. 6 anti-cytokeratin antibodies, CK8, 18, 19, CK7, CK8, CK14, CK19 and AE1/AE3 were used against the keratin proteins expressed from the human meibomian fluids. Using the immunoblotting (dot blot) technique, abnormal waxy meibomian fluids obtained from subjects diagnosed to have meibomian gland dysfunction (MGD) were compared to normal clear meibomian fluids. The results show that keratins are present in a higher concentration (10%) in the abnormal human meibomian excreta as compared to the normals. Even though the presence of protein markers for keratinization in the abnormal meibomian excreta were not shown, the increased presence of keratin proteins in the abnormal meibomian fluids suggests that, in MGD patients, hyperkeratinization of ductal epithelium may have taken place. More keratin proteins (possibly those of higher molecular weights) were produced in addition to the keratin proteins normally produced by the duct epithelium. The increased amount of keratin proteins in the abnormal meibomian fluids may be explained by the susceptibility of duct epithelium to undergo the process of hyperkeratinization as postulated by other researchers.


Ecotropic viral integration site 1, a novel oncogene in prostate cancer.
Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous cancer in men in the western world. Mutations in tumor suppressor genes and in oncogenes are important for PCa progression, whereas the role of stem cell proteins in prostate carcinogenesis is insufficiently examined. This study investigates the role of the transcriptional regulator Ecotropic Viral Integration site 1 (EVI1), known as an essential modulator of hematopoietic and leukemic stem cell biology, in prostate carcinogenesis. We show that in healthy prostatic tissue, EVI1 expression is confined to the prostate stem cell compartment located at the basal layer, as identified by the stem cell marker CD44. Instead, in a PCa progression cohort comprising 219 samples from patients with primary PCa, lymph node and distant metastases, EVI1 protein was heterogeneously distributed within samples and high expression is associated with tumor progression (P<0.001), suggesting EVI1 induction as a driver event. Functionally, short hairpin RNA-mediated knockdown of EVI1 inhibited proliferation, cell cycle progression, migratory capacity and anchorage-independent growth of human PCa cells, while enhancing their apoptosis sensitivity. Interestingly, modulation of EVI1 expression also strongly regulated stem cell properties (including expression of the stem cell marker SOX2) and in vivo tumor initiation capacity. Further emphasizing a functional correlation between EVI1 induction and tumor progression, upregulation of EVI1 expression was noted in experimentally derived docetaxel-resistant PCa cells. Importantly, knockdown of EVI1 in these cells restored sensitivity to docetaxel, in part by downregulating anti-apoptotic BCL2. Together, these data indicate EVI1 as a novel molecular regulator of PCa progression and therapy resistance that may control prostate carcinogenesis at the stem cell level.


Hydroalcoholic extract from Baccharis dracunculifolia recovers the gastric ulcerated tissue, and p-coumaric acid is a pivotal bioactive compound to this action.
Baccharis dracunculifolia is a medicinal plant native to southeastern Brazil and is the main botanical source used by bees (Apis mellifera) in the manufacture of green propolis and display similar gastroprotective action and chemical profile. This article reports the healing gastric ulcer activity of the hydroethanolic extract of B. dracunculifolia (HEBD) in an acetic acid-induced ulcer model. In addition to the extract, the isolated compounds ferulic acid, p-coumaric acid, caffeic acid, baccharin, and aromadendrin-4'-O-methyl ether were also assayed. HEBD at a dose of 300 mg/kg reduced the ulcerated area by 49.4% after treatment for 7 days, twice a day. Histological analyses revealed that the margins and base of the ulcer obtained significant regeneration, and periodic acid Schiff base staining showed a 78.2% increase in the mucin levels. The action on the enzymatic antioxidant system demonstrated an increased activity of superoxide dismutase and glutathione-S-transferase, in addition to raising glutathione reduced levels and myeloperoxidase activity. HEBD did not show cytotoxicity in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylterazole bromine test. In vitro, HEBD inhibited the H+ /K+ -ATPase enzyme and showed antioxidant activity in the 2,2 diphenyl-1-picryllydrazyl test. Regarding the isolated compounds, oral administration of p-coumaric acid (15 mg/kg) reduced the ulcerated area by 66.2%. The results suggest that HEBD recovers the gastric ulcerated tissue, raising mucus and antioxidant enzyme levels, and reducing the H+ /K+ -ATPase activity. In addition, the findings confirm that p-coumaric acid is a pivotal bioactive compound on the gastric healing effects elicited by HEBD. © 2019 BioFactors, 45(3):479-489, 2019.


Resolving front-back ambiguity with head rotation: The role of level dynamics.
Making small head movements facilitates spatial hearing by resolving front-back confusions, otherwise common in free field sound source localization. The changes in interaural time difference (ITD) in response to head rotation provide a robust front-back cue, but whether interaural level difference (ILD) can be used as a dynamic cue is not clear. Therefore, the purpose of the present study was to assess the usefulness of dynamic ILD as a localization cue. The results show that human listeners were capable of correctly indicating the front-back dimension of high-frequency sinusoids based on level dynamics in free field conditions, but only if a wide movement range was allowed (±40∘). When the free field conditions were replaced by simplistic headphone stimulation, front-back responses were in agreement with the simulated source directions even with relatively small movement ranges (±5∘), whenever monaural sound level and ILD changed monotonically in response to head rotation. In conclusion, human listeners can use level dynamics as a front-back localization cue when the dynamics are monotonic. However, in free field conditions and particularly for narrowband target signals, this is often not the case. Therefore, the primary limiting factor in the use of dynamic level cues resides in the acoustic domain behavior of the cue itself, rather than in potential processing limitations or strategies of the human auditory system.


Architectural roles of Cren7 in folding crenarchaeal chromatin filament.
Archaea have evolved various strategies in chromosomal organization. While histone homologues exist in most archaeal phyla, Cren7 is a chromatin protein conserved in the Crenarchaeota. Here, we show that Cren7 preferentially binds DNA with AT-rich sequences over that with GC-rich sequences with a binding size of 6~7 bp. Structural studies of Cren7 in complex with either an 18-bp or a 20-bp double-stranded DNA fragment reveal that Cren7 binds to the minor groove of DNA as monomers in a head-to-tail manner. The neighboring Cren7 monomers are located on the opposite sides of the DNA duplex, with each introducing a single-step sharp kink by intercalation of the hydrophobic side chain of Leu28, bending the DNA into an S-shape conformation. A structural model for the chromatin fiber folded by Cren7 was established and verified by the analysis of cross-linked Cren7-DNA complexes by atomic force microscopy. Our results suggest that Cren7 differs significantly from Sul7, another chromatin protein conserved among Sulfolobus species, in both DNA binding and deformation. These data shed significant light on the strategy of chromosomal DNA organization in crenarchaea.


Crystallization of the collagen-like polypeptide (PPG)10 aboard the International Space Station. 2. Comparison of crystal quality by X-ray diffraction.
Crystals of the collagen-like polypeptide (PPG)(10) were obtained within the Advanced Protein Crystallization Facility on board the International Space Station, during the STS-105/STS-108 mission. The duration of this mission was such to ensure that the crystallization process had reached its end. Crystals were grown both in the presence and in the absence of agarose gel, to compare the quality of the crystals obtained from these different environments. As a result, crystals grown in the absence of agarose on Earth as well as in microgravity showed X-ray diffraction up to 1.15 A. The intensity/sigma ratio was slightly higher for microgravity grown crystals. Crystals grown in agarose gel, both in microgravity and on ground, showed a comparable diffraction power, with a resolution limit of 1.45 A.


How to improve the effectiveness of transurethral resection in nonmuscle invasive bladder cancer?
The high rate of early recurrences in nonmuscle invasive bladder cancer is considered to be strongly related to the effectiveness of transurethral resection (TUR). The aim of this article is to review methods, currently available or in development, that aim at improving TUR, with an emphasis on studies over the past year. It has been shown that re-TUR diminishes staging error and improves local tumour control. However, instead of simply repeating the standard procedure, it makes more sense to improve the technique itself. Modifications in TUR equipment, such as bipolar resection and laser treatment, mainly have the potential to reduce complication rates and possibly thereby improve the effectiveness of TUR. New imaging techniques that can be used during TUR such as narrow-band imaging and optical coherence tomography seem promising, whereas photodynamic diagnosis has already proven its potential role in improving the effectiveness of TUR. The currently available techniques of re-TUR and photodynamic diagnosis have demonstrated potential to improve effectiveness of TUR and should be used in selected cases. New techniques such as narrow-band imaging and optical coherence tomography seem promising, but more evidence is needed before these methods can be implemented in daily practice.


Toward tunable band gap and tunable dirac point in bilayer graphene with molecular doping.
The bilayer graphene has attracted considerable attention for potential applications in future electronics and optoelectronics because of the feasibility to tune its band gap with a vertical displacement field to break the inversion symmetry. Surface chemical doping in bilayer graphene can induce an additional offset voltage to fundamentally affect the vertical displacement field and the band gap opening in bilayer graphene. In this study, we investigate the effect of chemical molecular doping on band gap opening in bilayer graphene devices with single or dual gate modulation. Chemical doping with benzyl viologen molecules modulates the displacement field to allow the opening of a transport band gap and the increase of the on/off ratio in the bilayer graphene transistors. Additionally, Fermi energy level in the opened gap can be rationally controlled by the amount of molecular doping to obtain bilayer graphene transistors with tunable Dirac points, which can be readily configured into functional devices, such as complementary inverters.


Acoustic signal characterization based on hidden Markov models with applications to geoacoustic inversions.
A probabilistic characterization scheme for acoustic signals with applications in acoustical oceanography is presented. This scheme aims at the definition of a set of stochastic observables that could characterize the signal. To this end, the signal is decomposed into several levels using the stationary wavelet packet transform. The extracted wavelet coefficients are then modeled by a hidden Markov model (HMM) with Gaussian emission distributions. The association of a signal with a representative HMM is performed utilizing the expectation-maximization algorithm. Eventually, the signal is characterized by the set of parameters that describe the HMM. The Kullback-Leibler divergence is employed as the similarity measure of two signals, comparing their corresponding HMMs. To validate the performance of the proposed characterization scheme, which is denoted as the probabilistic signal characterization scheme (PSCS), a simulated and a real experiment have been considered. The measured signal is characterized by the proposed PSCS method, and the model parameters of the seabed are estimated by means of an inversion procedure employing a genetic algorithm. The inversion results confirmed the reliability and efficiency of the proposed method when applied with typical signals used in applications of acoustical oceanography.


[Design features of an individual thrust plate prosthesis].
The purpose of the present study was to document the osteotomy plane for the thrust plate prosthesis, and to evaluate the question whether the geometry of the thrust plate itself correlates with the range of motion after implantation and whether the osteointegration area can be optimised. For the first part of the study, the two-dimensional geometry of the osteotomy was demonstrated in 12 computer-reconstructed femurs after performing a virtual cut at a CCD angle of 135 degrees. In the second part we constructed a prototype of an I-TPP with an optimised thrust plate and corpus geometry. In a final step, we documented the range of motion with computer-aided movement mapping. The results showed a wide variance in osteotomy geometry in the 12 femurs. With the I-TAP we were able to obtain a much better surface adaptation of the thrust plate. Movement mapping showed a much lower range of motion in the I-TPP implant.


Proliferation of gastric endocrine cells after vagotomy in the rat.
The effects of truncal vagotomy on gastrin cell and somatostatin cell populations and circulating hormone levels were studied in rats. Truncal vagotomy produced significant gastrin cell hyperplasia, with the largest increase in gastrin cell density observed 2 weeks postvagotomy. Gastrin cell hyperplasia was sustained up to 6 months. Both antral somatostatin cell and corpus somatostatin cell numbers demonstrated significant and sustained increases after vagotomy; the largest increases were observed 2 weeks after operation. Early elevation of circulating gastrin to 3 times the baseline level was followed by sustained, lower level hypergastrinemia. Plasma somatostatin levels were decreased 2 days after vagotomy, with a return to control values 2 weeks after operation. Gastrin cells and somatostatin cells are dynamic populations, capable of changes in both number and secretory function after truncal vagotomy.


Identification of a low specificity, oxygen, heme, and growth phase regulated DNA binding activity in Saccharomyces cerevisiae.
Cytochrome c oxidase from Saccharomyces cerevisiae is subject to intricate control at the level of transcription of the various genes encoding its subunits. Expression of the subunit VI encoding gene, COX6, is glucose repressed, growth phase induced, and dependent on oxygen and heme availability. An upstream activation region for COX6, UAS6, was found to contain a glucose responsive region, a heme dependent region (HDS1), and a binding site for the transcription factor, BAF1. BAF1 was the only factor observed to form a protein complex with UAS6 in vitro. However, we found that binding of BAF1 was unaffected by oxygen or heme regulation. In the present communication, we have identified a DNA binding activity that was growth phase induced and dependent on oxygen and heme availability. This highly regulated activity was detected by its discrete binding to the heme responsive site, HDS1, in UAS6. Nonetheless, it appears to have weak DNA binding specificity.


Detecting adaptive immunity: applications in transplantation monitoring.
In recent decades, continuous improvements in immunosuppressive therapy have led to a significant increase in kidney allograft survival. Despite innovative developments and improvements in immunosuppression, chronic allograft injury and late graft loss still remain major causes of morbidity and mortality. In clinical practice, long-term immunosuppression is adapted and fine-tuned according to drug levels, kidney function, and biopsy results. As an invasive procedure, indication biopsy still represents an indispensible diagnostic gold standard. However, in an effort to further improve outcomes on the basis of individualized treatment, there is an urgent need for noninvasive assays, as well as biomarkers, to more accurately monitor allogeneic responses and predict the risk of acute and chronic allograft rejection. This article discusses strategies for immune monitoring of T-cell responsiveness and humoral alloreactivity. Furthermore, new microarray and gene profiling data are highlighted, which may identify hyporesponsive transplant recipients who could benefit from a reduction or even withdrawal of immunosuppression. Finally, supplementary transplant risk assessment markers, such as soluble CD30 and urinary effector molecule analysis, are discussed as promising new tools. Recent developments and improvements in test principles to monitor and predict allograft immunity are encouraging and may herald the transition of present empiric immunosuppression to individualized immunosuppressive treatment. Nonetheless, before implementation of immune monitoring in routine clinical practice, there is still a need for prospective trials designed to clarify the actual diagnostic potential of individual test systems in a therapeutic context.


Cardiac and systemic rejuvenation after cardiosphere-derived cell therapy in senescent rats.
The aim is to assess the effects of CDCs on heart structure, function, gene expression, and systemic parameters in aged rats. Diastolic dysfunction is characteristic of aged hearts. Cardiosphere-derived cell (CDC) therapy has exhibited several favourable effects on heart structure and function in humans and in preclinical models; however, the effects of CDCs on aging have not been evaluated. We compared intra-cardiac injections of neonatal rat CDCs to vehicle (phosphate-buffered saline, PBS) in 21.8 ± 1.6 month-old rats (mean ± standard deviation; n = 23 total). Ten rats 4.1 ± 1.5 months of age comprised a young reference group. Blood, echocardiographic, haemodynamic and treadmill stress tests were performed at baseline in all animals, and 1 month after treatment in old animals. Histology and the transcriptome were assessed after terminal phenotyping. For in vitro studies, human heart progenitors from older donors, or cardiomyocytes from aged rats were exposed to human CDCs or exosomes secreted by CDCs (CDC-XO) from paediatric donors. Transcriptomic analysis revealed that CDCs, but not PBS, recapitulated a youthful pattern of gene expression in the hearts of old animals (85.5% of genes differentially expressed, P < 0.05). Telomeres in heart cells were longer in CDC-transplanted animals (P < 0.0001 vs. PBS). Cardiosphere-derived cells attenuated hypertrophy by echo (P < 0.01); histology confirmed decreases in cardiomyocyte area (P < 0.0001) and myocardial fibrosis (P < 0.05) vs. PBS. Cardiosphere-derived cell injection improved diastolic dysfunction [lower E/A (P < 0.01), E/E' (P = 0.05), end-diastolic pressure-volume relationship (P < 0.05) compared with baseline), and lowered serum brain natriuretic peptide (both P < 0.05 vs. PBS). In CDC-transplanted old rats, exercise capacity increased ∼20% (P < 0.05 vs. baseline), body weight decreased ∼30% less (P = 0.05 vs. PBS) and hair regrowth after shaving was more robust (P < 0.05 vs. PBS). Serum biomarkers of inflammation (IL-10, IL-1b, and IL-6) improved in the CDC group (P < 0.05 for each, all vs. PBS). Young CDCs secrete exosomes which increase telomerase activity, elongate telomere length, and reduce the number of senescent human heart cells in culture. Young CDCs rejuvenate old animals as gauged by cardiac gene expression, heart function, exercise capacity, and systemic biomarkers.


Impact of familial and hereditary prostate cancer on cancer specific survival after radical retropubic prostatectomy.
Men with a family history of prostate cancer are at higher risk for prostate cancer. There are conflicting data regarding the impact of hereditary forms of prostate cancer on long-term outcomes after radical prostatectomy. We examined the impact of familial and hereditary prostate cancer treatment in the prostate specific antigen era. Patients who underwent radical prostatectomy for prostate cancer from 1987 to 1997 were surveyed (3,560 responders) to determine the family history of prostate cancer. Patients were categorized as having familial prostate cancer if they had at least 1 first-degree relative with prostate cancer. Hereditary prostate cancer was defined as nuclear families with 3 cases of prostate cancer, families with prostate cancer in each of 3 generations and families with 2 men diagnosed before age 55 years. Sporadic prostate cancer was defined as patients with no family history. Clinical and pathological features, and long-term outcome measures, including biochemical recurrence-free, systemic progression-free and cancer specific survival, were compared among patients with familial, hereditary and sporadic prostate cancer. A total of 865 and 133 patients were categorized as having familial prostate cancer and hereditary prostate cancer, respectively. Preoperatively prostate specific antigen was higher in patients with hereditary prostate cancer than in the other 2 groups (p = 0.04). Ten-year biochemical progression-free, systemic progression-free and cancer specific survival were equivalent. Except for preoperative prostate specific antigen, clinicopathological features and long-term oncological outcomes are equivalent after radical prostatectomy in patients with familial, hereditary and sporadic prostate cancer.


Stellate Ganglion Block as a Diagnostic and Therapeutic Option in Chronic Pericardial Pain: A Case Report.
The underlying pathophysiology and treatment of chronic pericardial chest pain remains unclear. We describe a clinical case of a 38-year-old patient with chronic chest pain in the context of Marfan syndrome, status post-valve-sparing aortic root repair, and recurrent pericarditis. The patient suffers from chronic pericardial pain secondary to recurrent pericarditis refractory to pharmacotherapy. A left-sided stellate ganglion block (SGB) was performed for both diagnostic and therapeutic purposes. Postprocedure follow-up demonstrated significant analgesic benefit at 8 months after the procedure.


(99m)Tc hexamethyl-propylene-aminoxime single-photon emission computed tomography prediction of conversion from mild cognitive impairment to Alzheimer disease.
To examine the utility of single-photon emission computed tomography (SPECT) to predict conversion from mild cognitive impairment (MCI) to Alzheimer disease (AD). Longitudinal, prospective study. University-based memory disorders clinic. One hundred twenty seven patients with MCI and 59 healthy comparison subjects followed up for 1-9 years. Diagnostic evaluation, neuropsychological tests, social/cognitive function, olfactory identification, apolipoprotein E genotype, magnetic resonance imaging, and brain Tc hexamethyl-propylene-aminoxime SPECT scan with visual ratings, and region of interest (ROI) analyses were done. Visual ratings of SPECT temporal and parietal blood flow did not distinguish eventual MCI converters to AD (N = 31) from nonconverters (N = 96), but the global rating predicted conversion (41.9% sensitivity and 82.3% specificity, Fisher's exact test p = 0.013). Blood flow in each ROI was not predictive, but when dichotomized at the median value of the patients with MCI, low flow increased the hazard of conversion to AD for parietal (hazard ratio: 2.96, 95% confidence interval: 1.16-7.53, p = 0.023) and medial temporal regions (hazard ratio: 3.12, 95% confidence interval: 1.14-8.56, p = 0.027). In the 3-year follow-up sample, low parietal (p <0.05) and medial temporal (p <0.01) flow predicted conversion to AD, with or without controlling for age, Mini-Mental State Examination, and apolipoprotein E ε4 genotype. These measures lost significance when other strong predictors were included in logistic regression analyses: verbal memory, social/cognitive functioning, olfactory identification deficits, hippocampal, and entorhinal cortex volumes. SPECT visual ratings showed limited utility in predicting MCI conversion to AD. The modest predictive utility of quantified low parietal and medial temporal flow using SPECT may decrease when other stronger predictors are available.


[The end of antibiotics? Increasing bacterial resistance in global and Norwegian perspective].
During the last 20 years there has emerged a growing world-wide problem with regard to multidrug-resistant microbes. The most serious examples so far are vancomycin-resistant strains of Enterococcus faecium, totally resistant isolates of Mycobacterium tuberculosis and multiple-resistant Staphylococcus aureus and Streptococcus pneumoniae. With the exception of some few strains of methicillin-resistant S. aureus and vancomycin-resistant enterococci, such bacteria have not been found in Norway. In this article we discuss possible ways of preventing further selection and spread of multiple-resistant microbes. We stress the importance of infection control programmes and restrictive use of antibiotics.