Anti-viral immune response of allogeneic irradiation bone marrow chimeras: cytotoxic T cell responsiveness depends upon H-2 combination and infectious agent.
Allogeneic irradiation bone marrow chimeras C57BL/10 (H-2b) leads to B10.BR (H-2k) and B10.BR (H-2k) leads to C57BL/10 (H-2b) were raised under specific pathogen-free (SPF) conditions; they survived very well and were healthy under SPF for 3-4 months and subsequently, under conventional housing conditions, for 1 to 8 months. Their immune response against third-party alloantigens was comparable with that of controls. Anti-vaccinia virus responses were very low when compared with syngeneic control chimeras or unmanipulated control animals; if anti-vaccinia virus cytotoxic T cell reactivity was measurable, it was specific for the bone marrow donor, rather than the recipient thymic H-2 type. In contrast, the anti-LCMV (lymphocytic-choriomeningitis virus) response was excellent and comparable to that in controls for B10.BR (H-2k) leads to C57BL/10 (H-2b) chimeras, but was completely absent for C57BL/10 (H-2b) leads to 10.BR (H-2k) chimeras. LCMV-specific cytotoxic effector T cells from B10.BR leads to C57BL/10 chimeras were restricted entirely to recipient H-2b. In contrast to the asymmetric cytotoxic T cell response, both types of chimeras developed good primary footpad swelling reactions after local infection, which arose somewhat slower with LCMV than in control of chimeras. The capacity to control infection by Listeria monocytogenes was excellent for all controls and B10.BR leads to C57BL/10 chimeras but apparently absent in C57BL/10 leads to B10.BR chimeras. Differentiation of T cell restriction specificity for thymic H-2 is apparently most efficient, but it remains unclear whether the observed asymmetry reflects exaggerated immune response regulation in H-2-incompatible stem cell-thymus chimeras or differential cross-reactivities between restricting transplantation antigens.


Open Conversion with Explantation of Stent Grafts After Endovascular Aneurysm Repair for Abdominal Aortic Aneurysm.
Although endovascular aneurysm repair (EVAR) for abdominal aortic aneurysm (AAA) is widely used worldwide, the fact that it is associated with increased rates of reintervention has been considered a problem. This study aimed to analyze the outcomes of primary open AAA repair and open conversion with explantation of stent grafts after EVAR. In this retrospective study, we enrolled 1,120 patients (open repair, n = 664; EVAR, n = 456) who underwent AAA repair at Kobe University from 1999 to 2019. Of the 664 patients who underwent open repair, 121 (patients who underwent primary open repair (POR) as a concomitant procedure and patients with ruptured AAA) were excluded from the study. The outcomes of POR were compared with those of open conversion with explantation of stent grafts. Of the 543 patients who underwent open repair, 513 underwent POR and 30 underwent open conversion with explantation of stent grafts. The operation time for POR was significantly less than that for open conversion with explantation. During surgery, patients who underwent open conversion with explantation required significantly more transfusions of red cell concentrate, fresh frozen plasma, and platelet concentrate than those who underwent POR. Overall, 30 patients who underwent open conversion with explantation required a total of 48 reinterventions before surgery. Hospital mortality rates were 0.7% and 0% in the POR and open conversion with explantation groups, respectively (P = 0.62). Although overall survival at 5 years in the POR group was significantly better than that in the open conversion with explantation group (89.3 ± 1.7% vs. 79.5 ± 9.6%; P = 0.01), there were no significant differences between the 2 groups regarding the freedom from aortic event (hospital death, reintervention, and aortic death). According to the multivariate analysis, open conversion with explantation was not an independent risk factor for late death. There were 20 patients who were hesitant to undergo OCE, although we recommended OCE. In a subgroup analysis, the overall mean cost borne by patients who underwent EVAR was approximately 2.3 times higher compared with that borne by patients who underwent POR. Although demanding, both early and long-term outcomes of OCE have been favorable in our present study. OCE is highly recommended in patients with persistent sac enlargement after EVAR.


Zero-link polymerized hemoglobin (OxyVita®Hb) stabilizes the heme environment: potential for lowering vascular oxidative stress.
A study was conducted to compare the resistance to heme exposure between myoglobin, bovine hemoglobin, and OxyVita®Hb (zero-link-hemoglobin-polymer). The rate of release of heme-iron is related to the tissue oxidative stress that can elicit deleterious effects in clinical uses of blood substitutes. Experimental work has focused on the unfolding of hemoglobin molecules, resulting in heme loss, by using urea as a denaturant and analyzing the Soret spectral region. These unfolding studies provide evidence for both the structural integrity and redox stability of OxyVita®Hb and demonstrate that OxyVita®Hb does not readily unfold and its heme exposure/release is greatly reduced.


Image analysis of the chromatin organization in the nuclear domains of freeze fractured hepatocytes and lymphocytes.
The complex organization of the interphase nucleus can be analyzed, by way of thin sectioning and also freeze-fracture. This approach has previously been utilized in association with image analysis to quantitatively describe the organization of isolated rat liver nuclei and nuclear matrices. The main nuclear domains which, in section, present marked differences due to their electron-density, can be identified in replicas with more complex procedures, based on the quantitative evaluation of the number of particles per unit area and mainly by using image analysis. A quantitative analysis of the nuclear substructures has been performed by way of image analysis on in situ nuclei of freeze-fractured cells presenting marked differences in the heterochromatin quantity, such as hepatocytes and lymphocytes. The replicated nuclear particles have been classified according to their diameter and the obtained histograms have been quantitatively evaluated. The nuclear domains, heterochromatin, interchromatin, nucleolus, present characteristic ratios among the three main classes of particles; that is, ribonucleoproteins, solenoid filaments and solenoid fibre aggregates. The typical patterns of the nuclear domains can be further stressed by selecting a single class of particles and by examining its topographic localization. While interchromatin and nucleolar domains present a similar quantitative pattern in hepatocytes and lymphocytes, the heterochromatin of lymphocytes contains a significative higher percentage of solenoid aggregates than that of hepatocytes.


Cystic fibrosis bone disease treatment: Current knowledge and future directions.
Bone disease is a frequent complication in adolescents and adults with cystic fibrosis (CF). Early detection and monitoring of bone mineral density and multidisciplinary preventive care are necessary from childhood through adolescence to minimize CF-related bone disease (CFBD) in adult CF patients. Approaches to optimizing bone health include ensuring adequate nutrition, particularly intake of calcium and vitamins D and K, addressing other secondary causes of low bone density such as hypogonadism, encouraging weight bearing exercise, and avoiding bone toxic medications. Of the currently available anti-resorptive or anabolic osteoporosis medications, only bisphosphonates have been studied in individuals with CF. Future studies are needed to better understand the optimal approach for managing CFBD.


There and back again: An evolutionary perspective on long-distance coordination of plant growth and development.
Vascular plants, unlike bryophytes, have a strong root-shoot dichotomy in which the tissue systems are mutually interdependent; roots are completely dependent on shoots for photosynthetic sugars, and shoots are completely dependent on roots for water and mineral nutrients. Long-distance communication between shoot and root is therefore critical for the growth, development and survival of vascular plants, especially with regard to variable environmental conditions. However, this long-distance signalling does not appear an ancestral feature of land plants, and has likely arisen in vascular plants to service the radical alterations in body-plan seen in this taxon. In this review, we examine the defined hormonal root-to-shoot and shoot-to-root signalling pathways that coordinate the growth of vascular plants, with a particular view to understanding how these pathways may have evolved. We highlight the completely divergent roles of isopentenyl-adenine and trans-zeatin cytokinin species in long-distance signalling, and ask whether cytokinin can really be considered as a single class of hormones in the light of recent research. We also discuss the puzzlingly sparse evidence for auxin as a shoot-to-root signal, the evolutionary re-purposing of strigolactones and gibberellins as hormonal signals, and speculate on the possible role of sugars as long-distance signals. We conclude by discussing the 'design principles' of long-distance signalling in vascular plants.


Sequentially alternating pollutant scenarios of phenolic compounds in a continuous aerobic granular sludge reactor performing simultaneous partial nitritation and o-cresol biodegradation.
Industrial wastewater treatment plants must operate properly during the transient-state conditions often found in the industrial production. This study presents the performance of simultaneous partial nitritation and o-cresol biodegradation in a continuous aerobic granular reactor under sequentially alternating pollutant (SAP) scenarios. Three SAP scenarios were imposed during the operation of the granular reactor. In each one, a secondary recalcitrant compound (either p-nitrophenol (PNP), phenol or 2-chlorophenol (2CP)) were added for a short period of time to the regular influent containing only ammonium and o-cresol. Partial nitritation and o-cresol biodegradation were not inhibited by the presence of PNP or phenol and both compounds were fully biodegraded. On the contrary, the presence of 2CP strongly inhibited both processes within 2days. However, the reactor was recovered in a few days. These findings demonstrate that treatment of complex industrial wastewaters with variable influent composition is feasible in a continuous aerobic granular reactor.


The Calgary semantic decision project: concrete/abstract decision data for 10,000 English words.
Psycholinguistic research has been advanced by the development of word recognition megastudies. For instance, the English Lexicon Project (Balota et al., 2007) provides researchers with access to naming and lexical-decision latencies for over 40,000 words. In the present work, we extended the megastudy approach to a task that emphasizes semantic processing. Using a concrete/abstract semantic decision (i.e., does the word refer to something concrete or abstract?), we collected decision latencies and accuracy rates for 10,000 English words. The stimuli were concrete and abstract words selected from Brysbaert, Warriner, and Kuperman's (2013) comprehensive list of concreteness ratings. In total, 321 participants provided responses to 1,000 words each. Whereas semantic effects tend to be quite modest in naming and lexical decision studies, analyses of the concrete/abstract semantic decision responses show that a substantial proportion of variance can be explained by semantic variables. The item-level and trial-level data will be useful for other researchers interested in the semantic processing of concrete and abstract words.


Myelitis in inflammatory disorders associated with myelin oligodendrocyte glycoprotein antibody and aquaporin-4 antibody: A comparative study in Chinese Han patients.
Myelitis is an important clinical component of myelin oligodendrocyte glycoprotein antibody (MOG-ab)-associated disease (MOGAD) and aquaporin-4 antibody (AQP4-ab)-positive neuromyelitis optica spectrum disorder (NMOSD). The aim of this work was to evaluate the differentiating features of myelitis between the two diseases. Myelitis-related clinical and radiologic data from 130 patients with MOGAD and 125 patients with AQP4-ab-positive NMOSD were retrospectively reviewed and compared. A scoring model was established to differentiate MOG-ab-associated myelitis from AQP4-ab-associated myelitis. Overall, 29.2% (38/130) of patients with MOGAD and 66.4% (83/125) of patients with AQP4-ab-positive NMOSD had ever experienced myelitis. Compared with those with NMOSD, patients with MOGAD exhibited a lower frequency of myelitis, either during the first episode (p < 0.0001) or throughout the disease duration (p < 0.0001). Compared with AQP4-ab-associated myelitis, MOG-ab-associated myelitis manifested a higher male-to-female ratio (p < 0.0001), younger age at disease onset (p = 0.0004), more prodromic influenza-like symptoms (p = 0.030), more prodromic fever (p = 0.0003), more bowel and bladder dysfunction (p = 0.011), less painful tonic spasms (p < 0.0001), and lower Expanded Disability Status Scale scores after treatment (p < 0.0001). On magnetic resonance imaging, lower spinal cord lesions (p = 0.023), short-segment lesions (p = 0.021), conus involvement (p = 0.0001), and H sign (p < 0.0001) were more common in MOG-ab-associated myelitis. A scoring model with a cutoff value of 4 differentiated MOG-ab-associated myelitis from AQP4-ab-associated myelitis with a sensitivity of 87.9% and a specificity of 90.1%. Myelitis was less commonly observed in MOGAD and exhibited distinct features compared to those of AQP4-ab-positive NMOSD.


Some effects of an ACTH 4-9 analog (Org 2766) on human performance.
The effects (on human beings) of the ACTH analog Org 2766 were investigated for a range of performance tests: complicated serial reaction task, running memory span, verbal learning and non-verbal mental ability tests (closure flexibility and non-verbal abstraction). Subjective ratings on feelings were taken and heart rate was measured. Only the 30-min reaction task produced significant drug effects. In this task performance tends to deteriorate as a function of time on task. During the test period there is a gradual increase in the number of lapses in performance due to moments of inattention. This deterioration is counteracted by the ACTH 4-9 analog, a result also found in previous study using ACTH 4-10 (Org OI63). Thus ACTH fragments which are devoid of endocrine effects seem to have a beneficial effect on goal-directed motivation oriented on the requirements of the task.


Isolation and purification of flavonoid glycosides from Trollius ledebouri using high-speed counter-current chromatography by stepwise increasing the flow-rate of the mobile phase.
Three flavonoid glycosides including orientin, vitexin, quercetin-3-O-neohesperidoside and one unknown compound were isolated and purified by high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC from Trollius ledebouri Reichb., a traditional Chinese medicine. Preparative HSCCC with a two-phase solvent system composed of ethyl acetate-n-butanol-water (2:1:3, v/v/v) was successfully performed by increasing the flow-rate of the mobile phase from 1.5 to 2.5 ml/min after 190 min. Consequently, 95.8 mg orientin, 11.6 mg vitexin, 9.3 mg unknown compound with purities of over 97% and one partially purified peak fraction (contained quercetin-3-O-neohesperidoside at 85.1% purity) were obtained from 500 mg of the crude extract. Then the partially purified fraction was further purified by reversed-phase semi-preparative high-performance liquid chromatography. The structure identification of all pure fractions was carried out by UV, MS, 1H NMR and 13C NMR.


Volatile versus intravenous anaesthesia and perioperative neurocognitive disorders: anything to see here?
There is a potential differential effect of sevoflurane compared with propofol on postoperative delirium and other perioperative neurocognitive disorders. More generally, there are perhaps differences between volatile and intravenous anaesthetic agents in their possible impact on perioperative neurocognitive disorders. Strengths and limitations of a recent study in this journal and its contribution to our understanding of the impact of anaesthetic technique on perioperative neurocognitive disorders are discussed.


Usefulness of magnetic resonance imaging for evaluation of cardiovascular invasion: evaluation of sliding motion between thoracic mass and adjacent structures on cine MR images.
To determine the feasibility and usefulness of magnetic resonance imaging (MRI) for evaluating cardiovascular invasion of a thoracic mass by demonstrating the sliding motion between the mass and adjacent structures. Twenty-six patients (17 males and nine females, mean age = 49 years) were included in this study. They all had thoracic masses with equivocal cardiovascular invasion on chest CT scan and/or MRI that were surgically confirmed. The pathologic diagnoses were teratoma/thymic tumor (N = 12), lung cancer (N = 9), and other thoracic tumor (N = 5). Conventional T1/T2, contrast-enhanced, and breathheld ECG-gated cine MRI using a steady-state free precession (SSFP) technique was performed. The results were compared to the surgical reports. The cine MR images showed the presence of sliding motion in 39 structures in 20 patients, which were surgically confirmed as not being invaded, and 15 structures in six patients with the absence of sliding motion noted as tumor invasion. Therefore, the accuracy of the cine MR images was 94.4% (51/54) for evaluating cardiovascular invasion of a thoracic mass. MRI can provide additional information and improve the accuracy of preoperative staging for predicting cardiovascular invasion of a thoracic mass by evaluating the sliding motion.


Unequal cross-over is involved in human alpha satellite DNA rearrangements on a border of the satellite domain.
It can be invoked from the theory of tandem repeat homogenization that DNA on a satellite/non-satellite border may carry sequence marks of molecular processes basic to satellite evolution. We have sequenced a continuous 17-kb alpha satellite fragment bordering the non-satellite in human chromosome 21, which is devoid of higher-order repeated structure, contains multiple rearrangements, and exhibits higher divergence of monomers towards the border, indicating the lack of efficient homogenization. Remarkably, monomers have been found with mutually supplementary deletions matching each other as reciprocal products of unequal recombination, which provide evidence for unequal cross-over as a mechanism generating deletions in satellite DNA.


I feel you-monitoring environmental variables related to asthma in an integrated real-time frame.
The study of asthma and other complex diseases has proven to be a "moving target" for researchers due to its complex aetiology, difficulty in definition, and immeasurable environmental effects. A large number of studies regarding the contribution of both genetic and environmental factors often result in contradictory results, in part due to the highly heterogeneous nature of asthma. Recent literature has focused on the epigenetic signatures of asthma caused by environmental factors, highlighting the importance of environment. However, unlike the genetic techniques, environmental assessment still lacks accuracy. A plausible solution for this problem would be an individual-based environmental exposure assessment, relying on new technologies such as personal real-time environmental sensors. This could prove to enable the assessment of the whole environmental exposure-or exposome-matching in terms of precision the genome that is emphasized in most studies so far. In addition, the measurement of the whole array of biological molecules, in response to the environment action, could help understand the context of the disease. The current perspective comprises a beyond-genetics integrated vision of omics technology coupled with real-time environmental measures targeting to enhance our comprehension of the disease genesis.


Approximation of dose quality indicator values in multi-criteria optimized (MCO) volumetric modulated arc therapy (VMAT) treatment planning using trilinear dose interpolation.
To approximate dose-volume histogram (DVH) based quality indicators in volumetric modulated arc therapy (VMAT) planning using multi-criteria optimization (MCO) with a low number of composite optimization parameters. The solution space for VMAT optimization with a low number of composite optimization parameters is approximated by trilinear dose inter- polation and prediction of dose-volume-histogram (DVH) based plan quality indicator values. To assess the approximation quality a diverse dataset of 44 cranial and 18 spine patient geometries was chosen. Optimization results are governed by three composite parameters focusing on target-organ-at-risk- (OAR)-trade-off, overall healthy tissue sparing, and delivery/quality assurance complexity. 21,266 optimized dose distributions were pre-calculated and the numerical values for a choice of 10 DVH points, referred to as plan quality indicators, were stored to serve as ground truth. Using a subset of 8 and 27 pre-calculated optimization results, dose distributions for unknown parameter values were approximated by trilinear interpolation. The resulting quality indicator values were compared to the previously obtained exact solutions. The magnitude of the deviation between exact and approximated values varied largely with respect to patient geometry and the criterion under investigation. Approximation with 27 pre-calculated results yielded lower deviations than approximation with 8 results, at the cost of a higher pre-calculation workload. Solution space approximation via trilinear dose interpolation in VMAT treatment planning governed by composite optimization parameters is possible without further knowledge of the internal implementation of the underlying optimizer. Maximum average deviations between approxi- mation and actual values of characteristic dose quality indicators below 1% (cranial) and 8% (spine) allow for a quick qualitative assessment of the possible solution landscape.


Mutational analysis of SRY in XY females.
The Y chromosome located gene SRY (sex determining region Y gene) was identified in the search for the mammalian testis determining factor (TDF). Approaches for evaluating SRY as a candidate for TDF included the finding of mutations in SRY in the genomes of patients with failed testis development (XY females or 46,XY gonadal dysgenesis) and the production of female to male sex reversed mice transgenic for the mouse homologue of SRY, [Sry]. Since the initial use of XY females in the proof of SRY/TDF identity, many more patients have been analysed using different techniques and more mutations identified. A total of 11 mutations in SRY have now been described, all in the DNA-binding HMG-box region of the gene, and all in patients with apparently complete gonadal dysgenesis. Surprisingly, three familial SRY mutations have been identified, where the phenotype is either fertile male or sterile sex-reversed female. Estimates of the proportion of XY females mutant for SRY average at approximately 15%. Reasons for the low frequency of SRY mutations in XY sex reversal could be the presence of mutations in regions of SRY not yet discovered, the occurrence of mutations that give the same phenotype, perhaps in genes close to SRY in the testis determining pathway, or incorrect diagnosis of complete gonadal dysgenesis.


Mycophenolate mofetil: ten years' experience of a renal transplant unit.
Mycophenolate mofetil (MMF) use in renal transplantation has allowed a significant decrease in early acute rejection rates. We retrospectively evaluated the incidence of acute rejection episodes, renal function at the first year posttransplant, patient and graft survivals, cytomegalovirus (CMV) infection rate, influence of the degree of sensitization, and number of MHC antigen mismatches on graft survival in two groups of patients receiving either MMF or azathioprine. Group 1 included 149 patients receiving cyclosporine, MMF, and prednisolone; group 2 included 191 patients receiving cyclosporine, azathioprine, and prednisolone. The two groups did not differ in terms of age, sex, degree of sensitization (expressed as percentage of antibodies reactive to panel), MHC mismatch number, cold ischemia time, donor age, or anti-thymocyte globulin induction. In group 1 (MMF) there was a significant decrease in early acute rejection rate (19% vs 57%, P < .0001), longer graft survival at 10 years (92% vs 75%, P = .006), and higher rate of CMV infection (22% vs 12%, P = .004). Renal function at the first year posttransplant and patient survival during follow-up did not differ between the groups. The degree of sensitization influenced graft survival in group 2. The number of MHC mismatches did not influence graft survival in either group. With MMF, there was a significant reduction in early acute rejection rate, a significant increase in graft survival at 10-year follow-up, and diminished impact of the degree of sensitization on graft survival.


Diagnostic significance of denture complaints.
An inventory of denture complaints should be completed by patients who request replacement dentures prior to a diagnostic interview. Patient responses are often more spontaneous and unguarded and will reveal problems that might otherwise remain concealed. A more fruitful interview with the dentist will facilitate the information of a treatment plan that will satisfy that patient's need.


Comparative dietary sulfated metabolome analysis reveals unknown metabolic interactions of the gut microbiome and the human host.
The gut microbiome converts dietary compounds that are absorbed in the gastrointestinal tract and further metabolized by the human host. Sulfated metabolites are a major compound class derived from this co-metabolism and have been linked to disease development. In the present multidisciplinary study, we have investigated human urine samples from a dietary intervention study with 22 individuals collected before and after consumption of a polyphenol rich breakfast. These samples were analyzed utilizing our method combining enzymatic metabolite hydrolysis using an arylsulfatase and mass spectrometric metabolomics. Key to this study is the validation of 235 structurally diverse sulfated metabolites. We have identified 48 significantly upregulated metabolites upon dietary intervention including 11 previously unknown sulfated metabolites for this diet. We observed a large variation in subjects based on their potential to sulfate metabolites, which may be the foundation for classification of subjects as high and low sulfate metabolizers in future large cohort studies. The reported sulfatase-based method is a robust tool for the discovery of unknown microbiota-derived metabolites in human samples.


An integrated fecal microbiome and metabolome in the aged mice reveal anti-aging effects from the intestines and biochemical mechanism of FuFang zhenshu TiaoZhi(FTZ).
Fufang Zhenzhu Tiao Zhi (FTZ) capsule is a Chinese herbal preparation under the guidance of professor Guo Jiao's new theory of "Tiaogan Qishu Huazhuo" for disorders of glucose and lipid metabolism for more than twenty yares, which has been demonstrated to exhibit potential anti-aging effects such as regulation of glucose and lipid metabolisms and antiinflammatory and antioxidative effects. This study attempts to reveal the anti-intestinal aging effect and possible mechanism of FTZ. The mice were divided into three groups: the control group, model group and treatment group. The treatment group was given 1.0 g/kg body weight of FTZ extract administered by oral gavage once a day for 12 consecutive weeks. Age-related alterations such as HE staining of intestinal tissue morphology, mRNA levels of intestinal telomerase and inflammatory cytokines were observed Fecal samples were used for ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) and 16S rRNA gene sequencing analyses to reveal age-related metabolic perturbations and gut flora disorders to demonstrate the effects of FTZ. Emerged pathological morphology of intestinal tissues, upregulated relative expression level of gut inflammatory factors and decreased relative expression level of telomerase mRNA in aging mice illustrated characteristic senescent phenotypes in model group. FTZ treatment significantly lowered intestinal inflammation levels, enhanced telomerase activity, partially reversed the fecal metabolites abnormalities and restored the disorders of intestinal flora. Multiple potential metabolites were associated with linoleic acid, glycerophospholipid, α-linolenic acid, biosynthesis of unsaturated fatty acids and glycerolipid metabolisms. Several decreased beneficial butyrate-producing bacteria like S24-7, possible Alkaliphilus belonging to the Clostridia class and increased harmful bacteria such as potentially toxic metabolite hydrogen sulfide-producer Bilophila and Desulfovibrio, inflammation-mediator Mucispirillum were determined in present aging mice. These age-related poor alterations could be partially attenuated by FTZ treatment. The pathologic changes of intestinal senescence and the decrease of telomerase mRNA in elderly mice was observed. FTZ may sever as a novel delaying intestinal aging strategy via three pathways for anti-inflammatory, improving gut metabolites and gut flora. This study not only provided biological basis for the theory of treating different diseases with the same treatment in TCM, but also provided objective evidence for incorporating aging into the system of"glucose-lipid metabolism disease".


Structural heterogeneity of dermatan sulfate chains: correlation with heparin cofactor II activating properties.
To obtain dermatan sulfate (DS) with different structural characteristics and biological properties we isolated three groups of chains from a bovine mucosal DS preparation, differently iduronated and sulfated. The selected DS chains were characterized by their total charge values, electrophoretic mobility, susceptibility to Chondroitinase AC II treatment and disaccharide composition of Chondroitinase ABC digests. Besides the major IdoUA-->GalNac-4-SO4 sequences, two DS fractions (s-DS 1.75 M and f-DS 1.75 M) contained more than 10% disulfated disaccharides sequences and the third DS fraction (s-DS 1.5 M) contained only 4% disulfated disaccharides. Chondroitinase AC II treatment indicated that both the electrophoretically retarded forms (s-DS) are iduronic acid rich, as they were only minimally degraded to disaccharides/oligosaccharides. The ability of DS crude preparation to activate the heparin cofactor II (HCII) mediated inhibition of thrombin depends on the relative amount of highly active DS chains; this activity is related to the overall charge of DS chains and particularly with the content of IdoUA-2-SO4-->GalNAc-4-SO4 and UA-->GalNAc-4,6-di-SO4 disaccharides.


Separation of phenylthiohydantoin amino acids by capillary electrochromatography.
Capillary electrochromatography (CEC) was employed as a rapid and high-efficiency method for the isocratic separation of all 20 important phenylthiohydantoin (PTH) amino acids, the end products of Edman degradation during N-terminal protein sequencing. For this purpose, 75 microm ID fused-silica capillaries were packed with standard 3 microm Hypersil octadecyl silica (ODS) particles using a two-step column fabrication process, which represents a fast, reliable and efficient means of producing long-term stable columns. The influence of solvent composition, pH, type of buffer cation, buffer concentration, and temperature on retention behavior of PTH amino acids was investigated. Same-day and day-to-day reproducibility of the retention times (over a period of two months) were found to be better than 3%. When comparing this new technique with traditional reversed phase-high performance liquid chromatography (RP-HPLC) methods applied in automated protein sequenators, CEC shows essentially shorter separation times and superior resolution.


Functional simian immunodeficiency virus Gag-specific CD8+ intraepithelial lymphocytes in the mucosae of SIVmac251- or simian-human immunodeficiency virus KU2-infected macaques.
The vaginal and rectal mucosae are the first line of cellular immune defense to sexually transmitted human immunodeficiency virus type 1 (HIV-1) entry. Thus, intraepithelial lymphocytes (IELs) may be important in the immune response to HIV infection. Here we investigated whether functional IELs in mucosal compartments could be visualized by direct staining with a tetrameric complex specific for the simian immunodeficiency virus (SIV) immunodominant Gag epitope in either separated IEL cells or tissues of macaques infected with SIVmac251. Of the 15 Mamu-A*01-positive macaques studied here, eight were chronically infected with either SIVmac251 or simian-human immunodeficiency virus (SHIV) KU2 and the remaining seven were exposed mucosally to SIVmac251 and sacrificed within 48 h to assess the local immune response. Gag-specific CD8+ T-cells were found in separated IELs from the rectum, colon, jejunum, and vagina of most infected animals. Direct staining of tetramers also revealed their presence in intact tissue. These Gag-specific IELs expressed the activation marker CD69 and produced IFN-gamma, suggesting an active immune response in this locale.


Calcineurin inhibitor withdrawal and conversion to mycophenolate mofetil and steroids in cardiac transplant recipients with chronic renal failure: a word of caution.
Chronic renal failure (CRF) is a common complication of calcineurin inhibitor (CNI)-based immunosuppression following cardiac transplantation (HTx). The aim of this prospective study was to evaluate the impact of an immunosuppressive conversion from CNIs to mycophenolate mofetil (MMF) and steroids in cardiac transplant recipients with CRF on renal and cardiac graft function. Since 1999, 12 HTx recipients (10 men; 58 +/- 3.6 yr of age; 8.7 +/- 4.2 yr after HTx) with CNI-based immunosuppression and a calculated creatinine clearance (CreaCl) <50 mL/min were included. Most patients (10/12) were on cyclosporine and two patients were on tacrolimus prior inclusion. MMF was started with 0.5 g/d and adjusted according to the target trough levels (2-4 ng/mL). Prednisone dosage was 0.4 mg/kg. Subsequently, CNIs were completely withdrawn. Acute rejection episodes were excluded one and three months after conversion by endomyocardial biopsy and by echocardiography every three months thereafter. After a mean follow-up of 20 +/- 16 months, CreaCl improved significantly: pre-conversion vs. post-conversion: 32.8 +/- 12.2 mg/dL vs. 42.8 +/- 21.14 mg/dL, p = 0.03. However, four acute rejection episodes occurred and patients were reconverted to CNIs. Additionally, six patients had a new onset of graft vessel disease (GVD) one yr after conversion. As a result of these adverse events, the study was stopped after inclusion of only 12 of the scheduled 30 patients. Conversion to MMF and steroids after HTx improves renal function, but increases the risk for recurrent rejection and GVD. Therefore, MMF and steroids should only be considered in patients with a markedly low risk for rejection.


[Association between the beta 2 adrenergic receptor polymorphism and blood pressure in YueXi population].
To investigate the association between Arg16Gly polymorphism of beta(2)-adrenergic receptor (ADRB2) gene and blood pressure levels. A total of 487 hypertensive individuals were recruited from YueXi county of Anhui province. 672 patients' parents and siblings were also invited to take part in the study and used as genomic control. Blood pressure was measured and a standardized questionnaire regarding social-economic characteristics, general health status, occupational history and life-style and dietary factors was administered for each participant. The ADRB2 Arg16Gly polymorphism was genotyped by using a PCR-Restriction Fragment Length Polymorphism (RFLP) method. The association between the ADRB2 polymorphism and hypertension in hypertensive adults was determined by utilizing a family-based association test analysis. In this study population, carriers of the ADRB2 Arg16 allele had lower systolic (P < 0.01) and diastolic (P < 0.01) blood pressure, suggesting that the genetic effect on blood pressure was more likely to fit an additive model. Our results suggest a probable association between Arg16Gly polymorphism of ADRB2 gene and hypertension.


[Organ-preserving treatment as the most important option in the treatment of primary penile cancer].
The treatment of patients with penile cancer presents a significant challenge for therapists and patients alike. In this context, the focus must not only be on surgical techniques but rather on patient education and collaborative treatment planning. Consistent information about potential organ preservation and its consequences is essential. Following clinical examination and histological confirmation of the diagnosis, an organ-preserving and stage-adapted surgical procedure should be used in almost all cases. Intraoperatively, the aim should be to achieve tumour-free surgical margins through rapid pathological examination. Psycho-oncological support should be offered to all patients. Centralisation of treatment appears highly desirable for Germany, considering the excellent outcomes observed in English and Dutch centres. Until this has been achieved, following the valuable experience of the "Second Opinion Network for Testicular Tumours", the Second Opinion platform "eKonsil Urologie" (https://www.ekonsil.org) can be used as a reference.


Magnetic resonance imaging of the proximal tibial epiphysis is suitable for statements as to the question of majority: a validation study in forensic age diagnostics.
Determining majority plays a key role for forensic age diagnostics in living individuals. Recent data suggest that magnetic resonance imaging (MRI) of the proximal tibial epiphysis (PTE) may be a suitable alternative or at least an additional tool to clarify whether an individual has reached majority. However, the reference data situation is still sparse. Hence, the present dual center study retrospectively analyzed routine MRI of the knee in 413 cases (214 males and 199 females) of a Western Caucasian population aged between 12 and 25 years. MRI was performed at 1.5 and 3.0 T clinical scanners using T1- and T2-weighted sequences. The classification system by Vieth et al. (Eur Radiol 2018; 28:3255-3262) was applied for determining the ossification stages of the PTE. Intra-observer agreement was "very good" (κ = 0.931), and inter-observer agreement was "good" (κ = 0.798). Minimum ages above the age of 18 years were observed with the final stage (stage 6) in either sex (20.27 years in males and 18.55 years in females). The results are not in contradiction with the previous data and can be considered a strong and valuable support of the so far existing database. Therefore, the investigation of the PTE using routine MRI (either at 1.5 or 3.0 T) could be taken into consideration for application in forensic age estimation practice in near future.


Language access services for Latinos with limited English proficiency: lessons learned from Hablamos Juntos.
The Robert Wood Johnson Foundation funded Hablamos Juntos (HJ), a $10-million multiyear demonstration to improve access to health care for Latinos with limited English proficiency and to explore cost-effective ways for health care organizations to provide language access services. In this manuscript, the authors draw on their experiences in evaluating HJ, provide brief descriptions of innovative interventions, estimate operating costs, and synthesize lessons learned about implementation. A number of barriers and facilitators are documented. The experience of HJ grantees provides guidance for organizations contemplating similar efforts. In particular, it highlights the need for health care organizations to involve physicians in the design and adoption of language services.


Trap-tuning in afterglow perovskite crystals through alkali metal ion doping.
Herein, we report a trap-tuning strategy involving alkali metal ions of Li+ and K+, which enabled a fine tuning of afterglow traps at 250 K. Through Li+/K+ doping, a red afterglow from Cs2AgInCl6:Mn was activated at room temperature, which was extended up to 3600 s at low temperature. Thermoluminescence measurement revealed a shallow trap at ∼0.50 eV below the conduction band, which could be shifted up- and down-ward by dopant species.


Oxidative stress markers in pregnant women who snore and fetal outcome: a case control study.
To determine the levels of oxidative stress markers in pregnant women who snore and compare with non-snoring pregnant women. Fetal outcome of these 2 groups was also evaluated. Prospective, case control study. Some 40 pregnant women who snored and 43 non-snoring pregnant women were evaluated. The glutathione peroxidase (GSH-Px), malondialdehyde (MDA) and myeloperoxidase (MPO) levels of the 2 groups were studied. Infant birthweight, Apgar scores, and other indicators of fetal outcome were obtained. The mean level of GSH-Px was significantly lower in the pregnant women who snored (p=0.005), while the mean level of MDA was significantly higher in this group (p=0.005). Levels of MPO were comparable between the groups (p>0.05). The pregnant women who snored did not have infants with evidence of an increase in compromised outcome. Although the pregnant women who snored had high levels of MDA, they did not appear to be at increased risk for delivering infants with fetal compromise.


Changes in regional cytochrome oxidase activity in the fetal, newborn and adult ovine brainstem.
Metabolic activity of specific brain regions (e.g. brainstem respiratory centers) may increase during the physiologic adaptations at birth. Since regional activity of cytochrome oxidase is correlated with the level of oxidative metabolism, cytochrome oxidase histochemistry was used to investigate whether there are sustained changes in metabolic activity within specific nuclei of the ovine brainstem during the perinatal period and whether further changes occur in the adult. Histochemistry was performed on 10-microns-thick frozen sections of the perinatal (130 d fetus, 140 d fetus, 8 h newborn and 10 d newborn) and adult ovine brainstem (n = 3 at each age). Computer-assisted image analysis was performed on 20 brainstem regions. A general decreasing trend, interrupted by a tendency for a transient increase at 8 h after birth was observed in most regions analyzed. Statistically significant decreases (P < 0.05) in cytochrome oxidase levels between the perinatal age groups and the adult were found in 7 brainstem nuclei studied: ambiguus, cuneate, inferior olivary, reticularis lateralis, spinal trigeminal, parabrachial and superior olivary nuclei. Within the perinatal period, the nucleus gracilis was the only region to show statistically significant decreases in 140 d fetus and 8 h newborns in all nuclei analyzed, but this change was not statistically significant (P > 0.1). These results indicate that the dramatic changes in physiology and environment at birth do not result in a significant change in the metabolic capacity of brainstem nuclei in the immediate perinatal period. However, more gradual developmental changes are observed in specific brainstem nuclei suggesting a decrease in neuronal activity occurs in these areas during development in the sheep.


Toward a Deuterium Feather Isoscape for Sub-Saharan Africa: Progress, Challenges and the Path Ahead.
A key challenge to the application of continent-wide feather isoscapes for geographic assignment of migrant birds is the lack of ground-truthed samples. This is especially true for long-distance Palearctic-Afrotropical migrants. We used spatially-explicit information on the δ2H composition of archived feathers from Green-backed/Grey-backed Camaroptera, to create a feather δ2H isoscape for sub-Saharan Africa. We sampled from 34 out of 41 sub-Saharan countries, totaling 205 sampling localities. Feather samples were obtained from museum collections (n = 224, from 1950 to 2014) for δ2H assay. Region, altitude, annual rainfall and seasonal patterns in precipitation were revealed as relevant explanatory variables for spatial patterns in feather δ2H. Predicted feather δ2H values ranged from -4.0 ‰ to -63.3 ‰, with higher values observed in the Great Rift Valley and South Africa, and lower values in central Africa. Our feather isoscape differed from that modelled previously using a precipitation δ2H isoscape and an assumed feather-to-precipitation calibration, but the relatively low model goodness fit (F10,213 = 5.98, p<0.001, R2 = 0.18) suggests that other, non-controlled variables might be driving observed geographic patterns in feather δ2H values. Additional ground-truthing studies are therefore recommended to improve the accuracy of the African feather δ2H isoscape.


Symmetrical Diamidates as a Class of Phosphate Prodrugs to Deliver the 5'-Monophosphate Forms of Anticancer Nucleoside Analogues.
The application of phosphorodiamidate technology to pyrimidine and purine nucleosides with anticancer activity to potentially overcome the resistance mechanisms associated with parent nucleosides is reported. Sixteen symmetrical phosphorodiamidates were prepared from the natural amino acids l-alanine and glycine. All the compounds were evaluated for their cytotoxic activity against a wide panel of solid and leukaemic tumour cell lines. In addition, a carboxypeptidase Y assay was performed on a representative phosphorodiamidate in order to reveal the putative bioactivation pathway for the reported phosphorodiamidate-type prodrugs.


Transcriptomic analysis reveals responses to a polluted sediment in the Mediterranean copepod Acartia clausi.
Marine sediments are regarded as sinks for several classes of contaminants. Characterization and effects of sediments on marine biota now require a multidisciplinary approach, which includes chemical and ecotoxicological analyses and molecular biomarkers. Here, a gene expression study was performed to measure the response of adult females of the Mediterranean copepod Acartia clausi to elutriates of polluted sediments (containing high concentrations of polycyclic aromatic hydrocarbons, PAHs, and heavy metals) from an industrial area in the Southern Tyrrhenian Sea (Bagnoli-Coroglio). Functional annotation of the A. clausi transcriptome generated as reference here, showed a good quality of the assembly and great homology with other copepod and crustacean sequences in public databases. This is one of the few available transcriptomic resources for this widespread copepod species of great ecological relevance in temperate coastal areas. Differential expression analysis between females exposed to the elutriate and those in control seawater identified 1000 differentially expressed genes, of which 743 up- and 257 down-regulated. Within the up-regulated genes, the most represented functions were related to proteolysis (lysosomal protease, peptidase, cathepsin), response to stress and detoxification (heat-shock protein, superoxide dismutase, glutathione-S-transferase, cytochrome P450), and cytoskeleton structure (α- and β-tubulin). Down-regulated genes were mostly involved with ribosome structure (ribosomal proteins) and DNA binding (histone proteins, transcription factors). Overall, these results suggest that processes such as transcription, translation, protein degradation, metabolism of biomolecules, reproduction, and xenobiotic detoxification were altered in the copepod in response to polluted elutriates. In conclusion, our results contribute to gaining information on the transcriptomic responses of copepods to polluted sediments. They will also prompt the selection of genes of interest to be used as biomarkers of exposure to PAHs and heavy metals in molecular toxicology studies on copepods, and in general, in comparative functional genomic studies on marine zooplankton.


Cas12c-DETECTOR: A specific and sensitive Cas12c-based DNA detection platform.
The robust and precise nucleic acid detection platform enormously influences the clinical diagnosis of human and plant pathogens, drastically affecting disease pandemic control. CRISPR-based nucleic acid detection tools have been successfully applied for rapid and sensitive nucleic acid detection. However, the T-rich protospacer adjacent motif (PAM), specificity, and sensitivity of the CRISPR-based nucleic acid detection tools limited its wide application. We first developed a new Cas12c-based nucleic acid detection platform (Cas12c-DETECTOR), recognizing a 5'-TG PAM and showing high sensitivity and specificity on examined targets. Our results indicate that Cas12c-DETECTOR coupling with the optimized single-guide RNA (sgRNA) can be applied to specifically identity single nucleotide polymorphism (SNP). Moreover, combined with pre-amplification and lateral flow strips or the visual fluorescence detection method, Cas12c-DETECTOR can be used to diagnose human and plant pathogens in practice. Therefore, our findings illustrated that Cas12c-DETECTOR is a robust, sensitive, precise, and practiced nucleic acid detection platform.


Factors associated with acute cardiac injury and their effects on mortality in patients with COVID-19.
To determine the incidence of acute cardiac injury (ACI), the factors associated with ACI and the in-hospital mortality in patients with COVID-19, especially in severe patients. All consecutive in-patients with laboratory-confirmed COVID-19 from Tongji Hospital in Wuhan during February 1 and March 29, 2020 were included. The demographic, clinical characteristics, laboratory, radiological and treatment data were collected. Univariate and Firth logistic regression analyses were used to identify factors associated with ACI and in-hospital mortality, and Kaplan-Meier method was used to estimate cumulative in-hospital mortality. Among 1031 patients included, 215 (20.7%) had ACI and 501 (48.6%) were severe cases. Overall, 165 patients died; all were from the severe group, and 131 (79.39%) had ACI. ACI (OR = 2.34, P = 0.009), male gender (OR = 2.58, P = 0.001), oximeter oxygen saturation (OR = 0.90, P < 0.001), lactate dehydrogenase (OR = 3.26, P < 0.001), interleukin-6 (IL-6) (OR = 8.59, P < 0.001), high sensitivity C-reactive protein (hs-CRP) (OR = 3.29, P = 0.016), N-terminal pro brain natriuretic peptide (NT-proBNP) (OR = 2.94, P = 0.001) were independent risk factors for the in-hospital mortality in severe patients. The mortality was significantly increased among severe patients with elevated hs-CRP, IL-6, hs-cTnI, and/or NT-proBNP. Moreover, the mortality was significantly higher in patients with elevation of both hs-cTnI and NT proBNP than in those with elevation of either of them. ACI develops in a substantial proportion of patients with COVID-19, and is associated with the disease severity and in-hospital mortality. A combination of hs-cTnI and NT-proBNP is valuable in predicting the mortality.


Proteomic investigation on chronic bladder irritation in the rat.
Interstitial cystitis (IC) is a painful bladder syndrome associated with urinary frequency and urgency. Elusive cause of IC makes its diagnosis only possible by exclusion in many cases. In this study, we used proteomics for identifying disease-associated proteins in a rat model of chronic bladder irritation. Chronic irritation of the rat bladder was caused by a brief (90 seconds) intravesical instillation of 0.2 mL of 0.4 N HCl. Whole bladders were collected at different time points after treatment, snap frozen, and nuclear and cytosolic protein extracts were obtained. Samples were resolved in standard 2-dimensional (2D) gels stained with an improved Coomasie stain or by differential gel electrophoresis (DIGE). Differentially expressed spots were excised and identified by MALDI-TOF MS/MS. Histologic and Western blot analyses were also performed. Bladder morphology and histologic appearance of bladder sections after HCl treatment reflected hemorrhage, edema, epithelial denudation, detrusor mastocytosis, and eosinophilia. Proteomic analysis of irritated rat bladder revealed marked overexpression of 4 nuclear proteins and marked underexpression of 1 nuclear protein compared with normal rat bladders. Among these proteins, inflammation-associated calgranulin A (over) and smooth muscle protein-22/transgelin (under) showed opposed expression patterns after bladder irritation. Presence of mast cells and eosinophils and overexpression of calgranulin A confirm the inflammatory component of HCl-irritated bladder. Altered expression of nuclear proteins is of particular interest because of their possible role as a prognostic marker in inflammatory bladder disorders. However, more studies are needed before clinical application of these findings can be established.


Treatment of stage III breast cancer. A panel discussion.
A major question in oncology today concerns the most appropriate therapy for locally advanced breast cancer. Realistic goals include effective local treatment and a prolonged disease-free interval for these patients, most of whom have incurable disease. Here, our panel discusses the roles of surgery, radiation therapy, chemotherapy, and hormonal therapy, and the proper sequencing of these modalities.


Priming the brain to capitalize on metaplasticity in stroke rehabilitation.
Repetitive transcranial magnetic stimulation (rTMS) is emerging as a potentially valuable intervention to augment the effects of behavioral therapy for stroke. When used in conjunction with other therapies, rTMS embraces the concept of metaplasticity. Due to homeostatic mechanisms inherent to metaplasticity, interventions known to be in isolation to enhance excitability can interact when applied successively under certain timing conditions and produce enhanced or opposite effects. Similar to "muscular wisdom," with its self-protective mechanisms, there also appears to be "synaptic wisdom" in neural networks with homeostatic processes that prevent over- and under-excitability. These processes have implications for both enhancing and suppressing the excitability effects from behavioral therapy. The purpose of this article is to relate the concept of metaplasticity, as derived from studies in humans who are healthy, to stroke rehabilitation and consider how it can be leveraged to maximize stroke outcomes.


Clinical value of automated fibrin generation markers in patients with septic shock: a SepsiCoag ancillary study.
An ancillary analysis to the SepsiCoag multicentric prospective observational study on patients entering an intensive care unit with septic shock evaluated the prognostic potential of fibrin generation markers (FGMs) tested at inclusion in the study, on survival at day 30. After centralization of samples, three automated FGMs were compared: D-dimers (DDi), fibrin/fibrinogen degradation products (FDP) and fibrin monomers (FM). FM was the single FGM that was significantly higher in non-surviving patients, area under the receiver-operator characteristic curve (AUCROC ): 0·617, P < 0·0001. Significantly higher International Society on Thrombosis and Haemostasis Disseminated Intravascular Coagulation (ISTH DIC) scores were calculated in non-survivors using each of the three FGMs. A dose-effect relationship was observed between ISTH DIC scores and non-survival, with highest significance obtained using FM as the FGM. An overt DIC diagnosis using the ISTH DIC score calculated using FM was a predictor of non-survival at day 30, independently from overt DIC diagnosis based on scores calculated using FDP or DDi. The AUCROC values testing the ability of the ISTH DIC score to predict non-survival were 0·650, 0·624 and 0·602 using FM, DDi and FDP, respectively, as the FGM. In patients with septic shock, among the commercially-available automated assays, automated FM is the FGM best related with late prognosis.


A mode of incipient growth in chemically induced signet ring cell carcinoma of the canine stomach.
A 31-month-old female mongrel dog was orally administered with 50 mg or 100 mg of N-nitrosobutylurea (NBU) in gelatin-capsule 3 times per week for 19 months with interposing periods of complete suspension. Thirty-four foci of signet ring cell carcinoma were found in the antral region of the stomach. The majority of the foci (31 foci) were early cancer, and the remaining foci were invasive cancer. In addition to these lesions, there was "a single gland cancer" in which a row of cancer cells was confined to a single gland. The whole gland was composed of two cell layers; the inner layer facing the lumen was normal gastric cells and the outer layer was atypical or neoplastic cells underlaid by the basement membrane. Mitosis was frequently observed on the bottom of the gland. Atypical or neoplastic cells seemed to mature gradually through a process of upward migration with increase in cytoplasmic Alcian blue-PAS and HID-AB positive mucin. Some of the cells rich in mucin moved into the lamina propria. The other cells remained in the flow of the regular cell renewal system of the normal gastric cells and reached the top of the gland. This observation revealed a mode of incipient gastric cancer growth, which starts and spreads within a single gland, before it invades the surrounding lamina propria.


Densely arranged two-dimensional silver nanoparticle assemblies with optical uniformity over vast areas as excellent surface-enhanced Raman scattering substrates.
Dense two-dimensional assemblies of silver nanoparticles were fabricated over vast areas (~19 cm(2)) by utilizing a simple liquid-liquid interface assembly technique. The Raman signal for p-aminothiophenol immobilized on the assemblies was drastically enhanced by plasmon coupling-induced hot spots, and the enhanced signal showed high spatial reproducibility.


Benchmarking the mARC performance - treatment time and dosimetric linearity.
The mARC technique is a hybrid rotational IMRT modality operating in "burst mode". While it is generally assumed that it will be slower than VMAT, the real limits of operation have not been defined so far. We here present the first systematic study of the technical limits on mARC treatment. The following scenarios are considered: 18, 30, 36 or 45 arclets per rotation (spacing between 20° and 8°), flat and flattening-filter-free (FFF) energy, arclet width 4° or 2°, from 1 MU/arclet to 1000 MU/plan. All scenarios are irradiated, treatment times are measured and treatment parameters reported. Dose linearity was assessed by point dose measurements of the 18 arclet plans with 1-30 MU per arclet. Minimum treatment times (no MLC movement, few MUs) depend strongly on the number of arclets per rotation (1minute for 18 arclets to 1:50min for 45 arclets), and rise linearly with MU/arclets after a given cut-off value depending on scenario, arclet width and available maximum dose rate. MLC movement adds up to 2minutes of treatment time, but generally less (ca. 45seconds in realistic plans). The rules by which irradiation parameters are selected by the firmware can be partly discovered. The choice of dose rate is most clearly defined. For the flat 6 MV energy, the highest available dose rate (300 MU/min) is always applied. For FFF 7 MV dose rate is reduced for arclets with few MUs, so that an arclet is irradiated in no less than 0.3 s. Only for the case of 1 MU/arclet can this constraint not be met (the technical limit on the dose rate if 500 MU/min for FFF 7 MV). In this case, dosimetric linearity is reduced. In all other instances, deviations from linearity at low MU remain below 2%. Treatment times of down to 90seconds are technically achievable for treatment with FFF beams using up to 36 arclets per rotation (arclet spacing every 10°) for up to 900 MU/plan, comparable to VMAT treatment times. The values provided here are meant to serve as a reference for the design of mARC plans (choice of arclets spacing etc.) and as minimum times against which the performance of different treatment planning systems can be evaluated.


Overproduction of Exopolysaccharide Colanic Acid by Escherichia coli by Strain Engineering and Media Optimization.
Colanic acid (CA) is one of the major bacterial exopolysaccharides. Due to its biological activities, CA has a significant commercial value. However, the cultivation conditions have not been optimized for the large-scale production of CA. Here, we constructed a CA-overproducing Escherichia coli strain (ΔwaaF) and statistically optimized its culture media for maximum CA production. Glucose and tryptone were found the optimal carbon and nitrogen sources, respectively. Fractional factorial design indicated tryptone and Na2HPO4 as the critical nutrients for CA production. Through further optimization, we achieved a maximum CA production of 1910.0 mg/L, which is approximately 12-fold higher than the amount obtained using the non-optimized medium initially used. The predicted value of CA production was comparable with experimental value (2052.8 mg/L) under the optimized conditions. This study constitutes a successful demonstration of media optimization for increased CA production, and paves the way for future research for achieving large-scale CA production.


Intraspecific divergence in essential oil content, composition and genes expression patterns of monoterpene synthesis in Origanum vulgare subsp. vulgare and subsp. gracile under salinity stress.
Oregano (Origanum vulgare L.), one of the important medicinal plants in the world, has valuable pharmacological compounds with antimicrobial, antiviral, antioxidant, anti-inflammatory, antispasmodic, antiurolithic, antiproliferative and neuroprotective activities. Phenolic monoterpenes such as thymol and carvacrol with many medical importance are found in Oregano essential oil. The biosynthesis of these compounds is carried out through the methyl erythritol-4 phosphate (MEP) pathway. Environmental stresses such as salinity might improve the secondary metabolites in medicinal plants. The influence of salinity stress (0 (control), 25, 50 and 100 mM NaCl) on the essential oil content, composition and expression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), γ-terpinene synthase (Ovtps2) and cytochrome P450 monooxygenases (CYP71D180) genes involved in thymol and carvacrol biosynthesis, was investigated in two oregano subspecies (vulgare and gracile). Essential oil content was increased at low NaCl concentration (25 mM) compared with non-stress conditions, whereas it was decreased as salinity stress intensified (50 and 100 mM). Essential oil content was significantly higher in subsp. gracile than subsp. vulgare. The highest (0.20 mL pot-1) and lowest (0.06 mL pot-1) amount of essential oil yield was obtained in subsp. gracile at 25 and 100 mM NaCl, respectively. The content of carvacrol, as the main component of essential oil, decreased with increasing salinity level in subsp. gracile, but increased in subsp. vulgare. The highest expression of DXR, Ovtps2 and CYP71D180 genes was observed at 50 mM NaCl in subsp. vulgare. While, in subsp. gracile, the expression of the mentioned genes decreased with increasing salinity levels. A positive correlation was obtained between the expression of DXR, Ovtps2 and CYP71D180 genes with carvacrol content in both subspecies. On the other hand, a negative correlation was found between the expression of CYP71D180 and carvacrol content in subsp. gracile. The findings of this study demonstrated that both oregano subspecies can tolerate NaCl salinity up to 50 mM without significant reduction in essential oil yield. Also, moderate salinity stress (50 mM NaCl) in subsp. vulgare might increase the carvacrol content partly via increment the expression levels of DXR, Ovtps2 and CYP71D180 genes.


The perioral muscle continuum affects premaxillary development in Wistar rats.
Craniofacial deformities involve soft tissue and skeletal abnormalities. Facial bone growth is based on congenital defects and iatrogenic factors, in which muscle activity is important. Understanding the effects of muscle function on facial bone growth may help us in clinical treatment. Although there have been some studies, fewer have focused on the effects of perioral muscle continuity on maxillary development, which needs further research. In our study, mimic perioral muscle surgeries were performed in twenty 3-day Wistar rats, which were divided into four equal groups, including five untreated rats as control (Ctrl), five rats by unilateral perioral muscle incision (MI), five rats by unilateral perioral muscle incision combined with muscle stripping (MIMS) and five rats treated by unilateral perioral muscle incision combined with periosteal stripping (MIPS). After six weeks, skulls were imaged and measured by micro-CT scan and hematoxylin-eosin staining. Differences in the rats' premaxilla were analyzed with self-contrasted and group-control studies. Compared with Ctrl group, there were significant premaxillary developmental defects in the affected side of the rats in all three surgical groups. In the affected side, both the width and the length of the premaxilla were less than the unaffected side, particularly in MIMS and MIPS groups. Group-control study showed that the ratio of premaxillary length of affected side to unaffected side had significant differences between MI and MIMS. The conclusion was that complete perioral muscle continuity with intact muscle attachment on the premaxilla is the driving force for the premaxillary development.


Discovery of potent pyrrolo-pyrimidine and purine HDAC inhibitors for the treatment of advanced prostate cancer.
The development of drugs for the treatment of advanced prostate cancer (PCA) remains a challenging task. In this study we have designed, synthesized and tested twenty-nine novel HDAC inhibitors based on three different zinc binding groups (trifluoromethyloxadiazole, hydroxamic acid, and 2-mercaptoacetamide). These warheads were conveniently tethered to variously substituted phenyl linkers and decorated with differently substituted pyrrolo-pyrimidine and purine cap groups. Remarkably, most of the compounds showed nanomolar inhibitory activity against HDAC6. To provide structural insights into the Structure-Activity Relationships (SAR) of the investigated compounds, docking of representative inhibitors and molecular dynamics of HDAC6-inhibitor complexes were performed. Compounds of the trifluoromethyloxadiazole and hydroxamic acid series exhibited promising anti-proliferative activities, HDAC6 targeting in PCA cells, and in vitro tumor selectivity. Representative compounds of the two series were tested for solubility, cell permeability and metabolic stability, demonstrating favorable in vitro drug-like properties. The more interesting compounds were subjected to migration assays, which revealed that compound 13 and, to a lesser extent, compound 15 inhibited the invasive behaviour of androgen-sensitive and -insensitive advanced prostate cancer cells. Compound 13 was profiled against all HDACs and found to inhibit all members of class II HDACs (except for HDAC10) and to be selective with respect to class I and class IV HDACs. Overall, compound 13 combines potent inhibitory activity and class II selectivity with favorable drug-like properties, an excellent anti-proliferative activity and marked anti-migration properties on PCA cells, making it an excellent lead candidate for further optimization.


Synthesis of p-nitrophenyl 6-O-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-beta-D-galactopyranoside and p-nitrophenyl O-beta-D-galactopyranosyl-(1 linked to 3)-O-(2-acetamido-2-dexoy-beta-D-glucopyranosyl)-(1 linked to 6)-beta-D-galactopyranoside.
Condensation of 2-methyl-(3,4,6-tri-O-acetyl-1,2-dideoxy-alpha-D-glucopyrano)-[2',1':4,5]-2-oxazoline with p-nitrophenyl 2,3-di-O-acetyl-beta-D-galactopyranoside (4), followed by saponification of the resulting disaccharide derivative, produced p-nitrophenyl 6-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-beta-D-galactopyranoside as a crystalline compound. Reaction of 2-methyl-[4,6-di-O-acetyl-1,2-dideoxy-3-O-(2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl)-alpha-D-glucopyrano]-[2',1':4,5]-2-oxazoline with 4 in a similar reaction-sequence provided the title trisaccharide compound.


ATP-sensitive potassium channels and endogenous adenosine are involved in spinal antinociception produced by locus coeruleus stimulation.
It has been known that locus coeruleus (LC) stimulation suppresses nociceptive discharges of the thalamic parafascicular (PF) neurons through the spinally descending adrenergic terminals which inhibit the transmission of nociceptive signals in the spinal dorsal horn. This experimental model was used in the present study to analyze the detailed processes that happened in the dorsal horn following norepinephrine release by preemptive intrathecal (i.t.) administration of related drugs in lightly urethane-anesthetized rats. The results showed that: (1) LC stimulation significantly inhibited the noxiously-evoked discharges of PF neurons; (2) the LC stimulation-produced antinociception in PF neurons could be blocked either by i.t. glibenclamide, an ATP-sensitive potassium (K(+)(ATP)) channel blocker, or by i.t. aminophylline, an adenosine receptor antagonist; (3) nociceptive discharges of PF neurons were also suppressed both by i.t. 5 -N-ethylcarboxamido-adenosine (NECA, an adenosine receptor agonist) and by i.t. nicorandil (a K(+)(ATP) channel opener); and (4) i.t. aminophylline blocked the suppression of PF nociceptive discharges induced by i.t. nicorandil, while i.t. glibenclamide showed no effect on the suppression of nociceptive discharges induced by i.t. NECA. These results suggest that: (1) K(+)(ATP) channels and endogenous adenosine may be involved in the mediation of spinal antinociception induced by descending adrenergic fibers originating from the LC; and (2) the opening of K(+)(ATP) channels precedes the release of adenosine in the cascade of mediation.


The bovine genomic DNA sequence data reveal three IGHV subgroups, only one of which is functionally expressed.
A comprehensive analysis of cattle shotgun sequencing data reveals 36 immunoglobulin heavy chain variable genes. The previously described bovine subgroup IGHV1 contains 10 functional genes with a conserved promoter including the consensus octamer and several other transcription factor binding sites, intact exons and matching cDNA sequences. Subgroups IGHV2 and IGHV3 consist entirely of pseudogenes. Thus, the bovine germline IGHV repertoire is very limited. The IGHV genes are distributed in mammalian clans I and II, while no clan III genes were detected. Clan-specific PCR of genomic DNA from cattle, sheep, Eurasian elk, white-tailed deer, pig and dolphin indicates highly dynamic evolution of IGHV gene usage within Cetartiodactyla. The bovine germline IGHV repertoire was probably generated by recent duplications of an IGHV1-IGHV2 homology unit. Immunoglobulin heavy chain genes are largely incorrectly assembled in the current cattle genome versions Btau_4.2 and UMD_3.1. FISH experiments confirm an IGHV locus close to terminus of BTA21.


Epoxyeicosatrienoic acids, 20-hydroxyeicosatetraenoic acid, and renal microvascular function.
The development of pharmacological, genetic, and biochemical tools have allowed for detailed studies to determine the contribution of cytochrome P450 (CYP) metabolites of arachidonic acid to renal microvascular function. Renal microvessels can generate CYP hydroxylase metabolites including 20-hydroxyeicosatetraenoic acid (20-HETE) and CYP epoxygenase metabolites, epoxyeicosatrienoic acids (EETs). 20-HETE constricts afferent arterioles and contributes to renal blood flow autoregulation. EETs act as endothelium-dependent hyperpolarizing factors (EDHFs) on the renal microcirculation. 20-HETE inhibits whereas EETs activate renal microvascular smooth muscle cell large-conductance calcium-activated K(+) channels (KCa). Likewise, 20-HETE renal microvascular actions are pro-hypertensive and EET actions are anti-hypertensive. These findings in the renal microvasculature and those of others have provided impetus for the development of enzymatic inhibitors, agonists, and antagonists for 20-HETE and EETs to determine their potential therapeutic value. Initial genetic studies and experimental studies with soluble epoxide hydrolase inhibitors to increase EETs, EET analogs, and 20-HETE inhibitors have demonstrated improved renal microvascular function in hypertension. These findings have demonstrated the important contributions that 20-HETE and EETs play in the regulation of renal microvascular function.


Dissociation of c-fos from ODC expression and neuronal differentiation in a PC12 subline stably transfected with an inducible N-ras oncogene.
In order to develop a model system for investigating the role of ras genes in neuronal differentiation, a construct consisting of a mouse N-ras oncogene linked to a dexamethasone-inducible promoter was devised and transfected into a subline of the PC12 rat pheochromocytoma cell line. Clonal lines were isolated which extended neurite-like processes within one day of exposure to dexamethasone. N-ras had a strong antiproliferative effect on these cells. These effects were reversible after removing dexamethasone. Elevation of mRNA for ornithine decarboxylase (ODC) was detected 6-18 hours after induction of N-ras by dexamethasone. The effects of ras on cell division, differentiation and cell size were analogous, but not identical to the effects of NGF on PC12 cells. One NGF action, induction of c-fos mRNA did not occur in ras-induced cells indicating that c-fos induction is unnecessary for both neurite outgrowth and for subsequent induction of ODC mRNA. The ability of ras to induce ODC, a division promoting enzyme, may also be relevant to the transforming actions of ras oncogenes.


Ablate and Pace: Is There Still a Role?
Despite the development of newer drugs and procedures to improve rhythm control, there is still a place for ablation of the atrioventricular junction (AVJ) in the management of selected patients with AF who are refractory to medical therapy, to improve quality of life, prevent ventricular dysfunction, and to optimize cardiac resynchronization therapy. We review all aspects of the "ablate and pace" strategy, from its history to patient selection, technique, outcomes and applications, and identify the need for randomized clinical trials to address some of the remaining questions regarding its application in some groups of patients.


Distribution, metabolism, and excretion of [14C] glutaraldehyde.
Radiolabeled glutaraldehyde (GA) was infused into rats in order to determine its distribution between cellular and humoral fractions of the blood, its potential metabolism by RBC's, its rate of excretion, and the nature of its urinary products. This study demonstrated that [14C]GA was distributed between the RBC's and plasma at a ratio greater than 1. Although the absolute counts of both fractions dropped 80% over 3 days, the percentage bound or incorporated increased over time. Despite the extensive uptake by RBC's these cells were unable to metabolize GA to CO2. Urinary excretion of the radiolabel was rapid; the predominant form in the urine was less than 1 kDa in size. All evidence suggested that it was not native GA. We conclude that the RBC's can incorporate GA, but can not metabolize it completely to CO2. Nevertheless, much of the infused GA was rapidly converted to nonreactive metabolites and eliminated by the kidney.


Proton-pump inhibitors associated with decreased urinary citrate excretion.
Proton-pump inhibitors (PPIs) may increase the risk of kidney stone formation, but the mechanism has not been elucidated. There is a paucity of literature evaluating the effects of PPIs on urinary metabolites and urine pH. We performed a retrospective review of nephrolithiasis patients treated at our institution and compared patients who were taking PPIs to those who were not at the time of their 24-h urine collections. Hierarchical multivariate linear regression was used to evaluate the independent relationship between PPI use and urinary mineral composition. We identified 301 consecutive patients, 88 (29%) of whom were taking PPIs at the time of their 24-h urine collections. Patients taking PPIs were older and more likely to have medical comorbidities associated with metabolic syndrome such as hypertension, diabetes, and dyslipidemia (p < 0.01). Controlling for these factors, patients taking PPIs were found to have 12% lower 24-h urine citrate excretion (β = - 0.12, ΔF = 4.24, p = 0.04). There were no other differences in urinary mineral composition between the groups. Our findings suggest that patients who take PPIs regularly may be at risk for decreased urinary citrate excretion. The consequent decrease in urinary citrate may become clinically significant for patients with other predisposing factors for hypocitraturia.


Apomorphine SL (Uprima): preclinical and clinical experiences learned from the first central nervous system-acting ED drug.
An exclusive central site of action for the proerectile effect of apomorphine, including not only the brain but also the spinal cord, is supported by extensive experimental data. Assuming that the mechanisms of action of apomorphine are similar in humans and animal models, its use for the treatment of erectile dysfunction (ED) validates the emerging idea that erectile response could be enhanced by acting directly within the central nervous system (CNS). It also emphasized the key role of the dopaminergic system in the control of erection. As exemplified with the clinical development of apomorphine, targeting the CNS does not rule out the occurrence of undesirable side effects. Because the rare event of syncope induced by apomorphine is not well understood, further research should be conducted to explore its possible mechanisms. In clinical practice, however, approved doses of apomorphine SL are well tolerated. It is noteworthy that no modification of sexual desire was observed with apomorphine. Indeed, drugs acting within the CNS may more likely interact with sexual desire than peripherally acting drugs, and care should be taken to assess this point in the future. Although our knowledge of the control of penile erection by the CNS is restricted, there are many potential sites for CNS-acting ED drugs. New centrally acting therapy for ED should concentrate on receptor targets more specific to erectile command. Clinical efficacy of new centrally-acting compounds will assess the well-founded purpose of this rationalization.


The Curved Presplitting Technique in a Totally Edentulous Atrophic Mandible: A Surgical Case Report.
The management of horizontally fully edentulous atrophic ridges is a common problem in dental implantology. This case report describes an alternative modified two-stage presplitting technique. The patient was referred for an implant-supported rehabilitation of their edentulous mandible. CBCT scans showed a mean available bone width of about 3 mm. At the first stage, four linear corticotomies were performed using a piezoelectric surgical device. At the second surgical stage 4 weeks later, bone expansion was performed, and four implants were placed in the interforaminal area. The healing process was uneventful. No fractures of the buccal wall and no neurologic lesions were observed. Postoperative CBCT scans showed a mean bone width gain of about 3.7 mm. Implants were uncovered 6 months after the second surgery, and 1 month later, a fixed provisional screw-retained prosthesis was delivered. This approach could be used as a reconstructive technique that avoids using grafts and reduces treatment times, possible complications, postsurgical morbidity, and costs by exploiting the patient's native bone as much as possible. Considering the limitations of a case report, randomized controlled clinical trials are needed to confirm the results and validate this technique.


The synaptic pathology of schizophrenia: is aberrant neurodevelopment and plasticity to blame?
Synaptic pathology is a feature of the brain in schizophrenia, denoted by alterations in the expression of synaptic proteins. In the absence of data indicative of neurodegenerative processes, the neuropathological features of schizophrenia suggest that the major pathogenic process in the disorder is one of aberrant development. Molecular evidence in support of a neurodevelopmental origin of schizophrenia has come from studies examining the expression of key developmental genes. However, as many of these genes are also involved in synaptic plasticity, their altered expression in schizophrenia also suggests that the disorder may be one of aberrant synaptic plasticity. The aim of this review is to explore whether aberrant development and synaptic plasticity may underlie the synaptic pathology of schizophrenia. It does this in two ways. First, studies in schizophrenia of the expression of two synaptic genes important in synaptic remodeling and plasticity are reviewed, changes in which may be indicative of aberrant synaptic plasticity in the disorder. Second, the possible relationship between the expression of genes involved in development and plasticity with that of presynaptic proteins is examined. Such a relationship, in combination with their altered expression in schizophrenia, may indicate whether developmental and plasticity-related processes may contribute to the synaptic pathology of the disorder. A brief discussion on the possible origins of the synaptic pathology of schizophrenia, and possible future studies, concludes the review.


Structural Characterization and Anticomplement Activity of an Acidic Heteropolysaccharide from Lysimachia christinae Hance.
A novel acidic heteropolysaccharide (LCP-90-1) was isolated and purified from a traditional "heat-clearing" Chinese medicine, Lysimachia christinae Hance. LCP-90-1 (Mw, 20.65 kDa) was composed of Man, Rha, GlcA, Glc, Gal, and Ara, with relative molar ratios of 1.00: 3.00: 11.62: 1.31: 1.64: 5.24. The backbone consisted of 1,4-α-D-GlcpA, 1,4-α-D-Glcp, 1,4-β-L-Rhap, and 1,3,5-α-L-Araf, with three branches of β-D-Galp-(1 → 4)-β-L-Rhap-(1→, α-L-Araf-(1→ and α-D-Manp-(1→ attached to the C-5 position of 1,3,5-α-L-Araf. LCP-90-1 exhibited potent anticomplement activity (CH50: 135.01 ± 0.68 µg/mL) in vitro, which was significantly enhanced with increased glucuronic acid (GlcA) content in its degradation production (LCP-90-1-A, CH50: 28.26 ± 0.39 µg/mL). However, both LCP-90-1 and LCP90-1-A were inactivated after reduction or complete acid hydrolysis. These observations indicated the important role of GlcA in LCP-90-1 and associated derivatives with respect to anticomplement activity. Similarly, compared with LCP-90-1, the antioxidant activity of LCP-90-1-A was also enhanced. Thus, polysaccharides with a high content of GlcA might be important and effective substances of L. christinae.


A phase I/II study to evaluate the effect of fractionated hemibody irradiation in the treatment of osseous metastases--RTOG 88-22.
The present study was initiated to determine the maximum tolerated total dose that can be delivered by fractionated hemibody irradiation (HBI), as defined by the acute hematological and nonhematological toxicity. Although it was designed as a dose searching trial, the influence of higher doses on occult and overt disease were considered equally important. The study was not designed to evaluate pain relief. The results were compared to Radiation Therapy Oncology Group (RTOG) 82-06, which employed single high-dose HBI, to determine if either single or fractionated HBI is more effective in controlling occult or overt disease. A total of 144 patients were entered from September 1989 to April 1993. Only patients with a single symptomatic bone metastases from either prostate or breast cancer primaries and a KPS > or = 60 were eligible. All patients initially received 30.0 Gy in 10 fractions to the symptomatic area followed by HBI in 2.50 Gy fractions to one of five arms: I-10.0 Gy (37 patients); II-12.5 Gy (23 patients); III-15.0 Gy (18 patients); IV-17.5 Gy (40 patients), and V-20.0 Gy (26 patients). A dose limiting toxicity was defined as an observed toxicity of > or = Grade 3 lasting more than 30 days postcompletion of HBI. If three or more dose-limiting toxicities occurred at any dose level, the previous dose was considered as the maximum tolerable dose. Thirty-six of 142 patients experienced > or = Grade 3 hematological toxicity at some time following HBI. The distribution of dose-limiting hematological toxicity in each arm was: I-two patients; II-one patients; III-zero patients; IV-one patient; and V-three patients. The major nonhematological toxicity was gastrointestinal and occurred in 10 patients. None were dose limiting. At 12 months from the initiation of treatment, the percent of patients with new disease were: Arms I-19%; II-9%; III-17%; IV-19%; V-13%; the percent of patients requiring additional treatment in the hemibody field were: Arms I-36%; II-30%; III-33%; IV-32%; and V-19%. When compared to single high-dose HBI the estimated reduction in the failure rate was 36% after fractionated HBI which potentially represents a modest improvement. The maximum tolerated dose of fractionated (2.50 Gy) HBI was found to be 17.5 Gy. The major dose limiting toxicity was hematological (thromboleukopenia). There was not a significant dose response effect on occult disease (appearance of new disease) or in the requirement for additional treatment, although certain trends were noted for the higher doses. When only patients completing assigned HBI from RTOG 82-06 and 88-22 were compared, there was no difference in the time to new disease or additional treatment in the treated field. Based on the investigative parameters of this study, single high-dose HBI was as effective as fractionated HBI. The incorporation of cytokines, to ameliorate hematological toxicity, should allow for the delivery of higher doses of fractionated HBI and sequential HBI as a means of delivering systemic irradiation.


Depressed myocardial contractility and normal ejection performance after aortic valve replacement in patients with aortic regurgitation.
Left ventricular cineangiography and pressure measurement were performed in 33 patients with chronic aortic regurgitation before and after aortic valve replacement and in 25 normal control subjects. Stress and volume were analyzed. The 33 patients were divided into three groups based on preoperative end-systolic volume index: 15 with an index of less than 100 ml/m2 (group I), 12 with an index of 100 to 200 ml/m2 (group II), and six with an index of more than 200 ml/m2 (group III). Afterload estimated as end-systolic stress, which was significantly elevated in all groups before operation, returned to normal postoperatively. Systolic pump function improved postoperatively in all groups; group I and group II showing the most improvement with group III still having an abnormal ejection fraction. The ratio of end-systolic stress to end-systolic volume index, which is an index of contractile state that is relatively independent of preload and afterload, was abnormal in each group preoperatively. Postoperatively, the ratio improved in each group (from 2.8 +/- 0.5 to 4.8 +/- 0.9 for group I, from 1.4 +/- 0.3 to 3.2 +/- 0.6 for group II, and from 0.8 +/- 0.2 to 2.6 +/- 0.8 for group III, compared with 5.9 +/- 1.0 for control subjects). Group I had normal values whereas group II and group III had subnormal ratios, suggesting a depressed contractility. All 15 patients in group I had values that fell within the 95% confidence limits of the linear inverse relationship between ejection fraction and end-systolic stress for control subjects. Nine of the 12 patients in group II and all six in group III had values that fell below the 95% confidence limits of the normal ejection fraction-end-systolic stress relationship. Aortic valve replacement may reduce afterload and improve systolic pump function in many patients with aortic regurgitation. However, there is a subgroup of patients who, despite normal or near-normal pump performance as assessed by ejection fraction, have depressed myocardial contractility characterized by an abnormal relationship between end-systolic stress and either ejection fraction or end-systolic volume.


[Susac syndrome: variable onset modes and disease courses: two case reports].
Susac syndrome is a rare microangiopathy of unknown etiology, which involves the clinical triad of encephalopathy, visual loss, and hearing loss. Several onset and progression patterns are possible. Case 1: a 43-year-old woman developed subacute encephalopathy, which had not been diagnosed and had been evolving for 2 months, as well as left sensorineural hearing loss. The fundus exam found central artery branch occlusion in the left retina. The treatment was based on corticosteroids combined with cyclophosphamide and immunoglobulins. Angiographic monitoring revealed persistent asymptomatic arterial alterations despite positive neurological progression. Case 2: a 27-year-old woman presented visual loss in the right eye after recurrent neurological episodes. The triad was completed by deafness in the right ear. Treatment with corticosteroids led to favorable neurological progression and stabilized the ophthalmologic symptoms. This syndrome preferentially affects young women. The nearly constant neurological symptoms can differ. Branch occlusions are frequently bilateral and often come with the appearance of vasculitis. Deafness is bilateral, asymmetrical, and of endocochlear origin. Brain MRI shows lesions of the corpus callosum that are distinctive of the syndrome. The disease mainly evolves in a monocyclic way, self-limited in time, and it rarely becomes chronic. Treatment, which has not been codified to date, is based on corticosteroids and, in severe cases, immunosuppressive drugs. Other therapies have not proved to be effective. The diagnosis is based on the triad of neurological, ophthalmic, and ENT damage, but sometimes it can be difficult to formulate because of the chronology of symptom onset. Neurological damage, the first manifestation, will help make therapeutic decisions.


Spatial and temporal distribution of Oct-4 and acetylated H4K5 in rabbit embryos.
Rabbit is a unique species to study human embryology; however, there are limited reports on the key transcription factors and epigenetic events of rabbit embryos. This study examined the Oct-4 and acetylated H4K5 (H4K5ac) patterns in rabbit embryos using immunochemistry staining. The average intensity of the Oct-4 signal in the nuclei of the whole embryo spiked upon fertilization, then decreased until the 8-cell stage and increased afterwards until the compact morula (CM) stage. It decreased thereafter from the CM stage to the early blastocyst (EB) stage, with a minimum at the expanded blastocyst (EXPB) stage and came back to a level similar to that of the CM-stage embryos in the hatching blastocysts (HB). The Oct-4 signal was observed in both the inner cell mass (ICM) and the trophectoderm (TE) cells of blastocysts. The average H4K5ac signal intensity of the whole embryo increased upon fertilization, started to decrease at the 4-cell stage, reached a minimum at the 8-cell stage, increased again at the EXPB stage and peaked at the HB stage. While TE cells maintained similar levels of H4K5ac throughout the blastocyst stages, ICM cells of HB showed higher levels of H4K5ac than those of EB and EXPB. Understanding key genetic and epigenetic events during early embryo development will help to identify factors contributing to embryo losses and consequently improve embryo survival rates. As a preferred laboratory species for many human disease studies such as atherosclerosis, rabbit is also a pioneer species in the development of several embryo biotechnologies, such as IVF, transgenesis, animal cloning, embryo cryopreservation and embryonic stem cells. However, there are limited reports on key transcription factors and epigenetic events of rabbit embryos. In the present study, we documented the temporal and spatial distribution of Oct-4 protein and H4K5 acetylation during early embryo development using the immunostaining approach. We also compared the patterns of these two important biomarkers between the inner cell mass (ICM) and the trophectoderm (TE) cells in blastocyst-stage embryos. Our findings suggest that a combination of Oct-4, H4K5ac and possibly other biomarkers such as Cdx-2 is needed to accurately identify different lineages of cells in morula and blastocyst stage rabbit embryos. Importantly, we revealed a novel wave of Oct-4 intensity change in the ICM cells of rabbit blastocysts. The signal was high at the early blastocyst stage, reached a minimum at the expanded blastocyst stage and returned to a high level at the hatching blastocyst stage. We hypothesize that the signal may have reflected the regulation of Oct-4 through enhancer switching and therefore may be related to cell lineage formation in rabbit embryos. These findings enrich our understanding on key genetic and epigenetic programming events during early embryo development in rabbits.


Effects of Eimeria maxima and Clostridium perfringens infections on cecal microbial composition and the possible correlation with body weight gain in broiler chickens.
With the voluntary and regulatory withdrawal of antibiotic growth promoters from animal feed, coccidiosis and necrotic enteritis (NE) emerge as the top two enteric poultry infectious diseases responsible for major economic loss worldwide. The objective of this study was to investigate the correlation between the cecal microbiota compositions with the growth trait after coccidiosis and NE. In this study, the effects of Eimeria maxima and/or Clostridium perfringens infections on the microbial composition and potential correlation with the body weight gain were investigated in broiler chickens using 16S rRNA gene sequencing. E. maxima and C. perfringens coinfection successfully induced NE with its typical gut lesions and significant reductions in the percentage of relative body weight gain (RBWG%). The NE challenge model did not affect cecal microbial diversity, but influenced the cecal microbial composition. KEGG enzymes in microbiota were significantly altered in abundance following dual infections. Furthermore, significant correlations between cecal microbiota modules and RBWG% were identified in the sham control, E. maxima or C. perfringens infected groups. Understanding of host-microbiota interaction in NE would enhance the development of antibiotics-independent strategies to reduce the harmful effect of NE on the gut microbiota structure, and improve the gut health and poultry production.


Tetraamine-modified octreotide and octreotate: labeling with 99mTc and preclinical comparison in AR4-2J cells and AR4-2J tumor-bearing mice.
Two somatostatin analogues, [(99m)Tc]Demotide and [(99m)Tc]Demotate 4, were compared with [(99m)Tc]Demotate 1, a previously reported somatostatin receptor subtype 2 (sst(2)) targeting tracer. Conjugates were prepared by coupling an open-chain tetraamine chelator to D-Phe(1) of [Tyr(3)]-octreotide or [Tyr(3)]-octreotate, respectively, via a p-benzylaminodiglycolic acid spacer adopting solid-phase peptide synthesis techniques. Peptide conjugates were collected in a highly pure form after chromatographic purification. Eventually, [(99m)Tc]Demotide and [(99m)Tc]Demotate 4 were obtained in approximately 1 Ci/micromol specific activity and >96% purity after labeling under alkaline conditions. Demotide and Demotate 4 exhibited similar high binding affinities for the sst(2) expressed in AR4-2J cells with IC(50) values 0.16 and 0.10 nM, respectively. The (radio)metallated analogues [(99m)Tc]Demotide and [(99m)Tc]Demotate 4 showed equally high affinities to the sst(2) during saturation binding assays in AR4-2J cell membranes (K(d)s 0.08 and 0.07 nM, respectively). During incubation at 37 degrees C with AR4-2J cells, the radiopeptides internalized effectively via a receptor-mediated process, with [(99m)Tc]Demotate 4 exhibiting a faster internalization rate than [(99m)Tc]Demotide. After injection in athymic mice bearing sst(2)-expressing AR4-2J tumors, the radiotracers showed high and specific uptake in the tumor (>25%ID/g at 1 h) and in the sst(2)-positive organs. However, both [(99m)Tc]Demotide and [(99m)Tc]Demotate 4 showed unfavorably higher background activity, especially in the abdomen, in comparison to [(99m)Tc]Demotate 1 and are, therefore, less suited than [(99m)Tc]Demotate 1 for sst(2)-targeted tumor imaging in man.


Prevalence of psychological disorders in the Athens area. Prediction of causal factors.
A two-phase cross-sectional study on the prevalence of psychological disorders was carried out in a sample of 1,574 adult male and female residents of two boroughs in the greater Athens area. The analysis focused on the detection of any possible effects (risk factors) that personal data (such as sociodemographic profile, use of psychiatric services, medication and experience of stressful life events) may have on mental health; information about the latter was assessed by Langner's scale. The findings are that all the independent variables investigated affect the respondents' psychological status, although interactive effects between variables predicted different patterns of psychopathological symptom formation.


An evaluation of Na+, C1- and pH ion-specific electrodes in the study of the electrolyte contents of epidermal transudate and sweat.
Ion-specific electrode detectors have been used to measure electrolytes in various biological fluids, including sweat. As part of a study of skin barrier function, we have used such electrodes to detect the presence of sodium, chloride and hydrogen (pH) in the transepidermal water loss. This paper describes the effects of cellophane tape stripping (of human forearm epidermis in vivo) on surface electrolyte concentrations, in parallel with observations on transepidermal water loss and galvanic skin resistance.