Structural basis for the recognition of superantigen streptococcal pyrogenic exotoxin A (SpeA1) by MHC class II molecules and T-cell receptors.
Streptococcal pyrogenic exotoxin A (SpeA) is a superantigen produced by Streptococcus pyogenes and is associated with severe infections characterized by rash, hypotension, multiorgan failure and a high mortality rate. In this study, an allelic form of this toxin, SpeA1, was crystallized with four molecules in the crystallographic asymmetric unit and its crystal structure was determined at 2.6 A resolution. The crystallographic R-factor was 19.4% (33 497 reflections) for 7031 protein atoms and 88 water molecules. The overall structure of SpeA1 is considerably similar to that of other prototype microbial superantigens, either of staphylococcal or streptococcal origin, but has greatest similarity to staphylococcal enterotoxin C (SEC). Based on structural and mutagenesis data, we have mapped several important residues on the toxin molecule, which are involved in the recognition of major histocompatibility complex (MHC) class II molecules and T-cell receptors. Also, the toxin appears to possess a potential zinc-binding site which may have implications in binding to particular MHC class II molecules. Finally, we propose models for SpeA1-MHC class II and SpeA1-T-cell receptor association and the relevance of this phenomenon to the superantigenic action of this toxin is considered.


[Effect of RNAi silencing of theWT1 gene on the apoptosis of human hepatocellular carcinoma cell hepG2].
To inhibit the expression of WT1 gene in HepG2 cells by siRNA, to study the effect of WT1 on the proliferation and apoptosis. HepG2 cells was transfected with WT1-siRNA by liposomes. The protein expression of WT1 in HepG2 cells was detected by immunocytochemistry to detect the effect of WT1 gene silencing. Tumor cells growth was detected by MTT assay. Morphology of apoptotic cells was observed by electron microscope and fluorescence. HepG2 cells was transfected with WT1-siRNA by liposomes, and the expression of WT1 were inhibited signifycantly. The transfection rate was over 70%.Silencing of WT1 by siRNA can inhibit cell proliferation and promote apoptosis. Sequence-specific siRNA targeting to WT1 can inhibit WT1 gene expression significantly. The decreesed expression of WT1 gene inhibits cell proliferation and promotes apoptosis of hepG2 cells.


Protein kinase C regulates proliferation of mast cells and the expression of the mRNAs of fos and jun proto-oncogenes during activation by IgE-Ag or calcium ionophore A23187.
Short-term stimulation (up to 16 hours) of interleukin-3 (IL-3)-dependent mouse bone marrow-derived mast cells, Abelson transformed mouse liver-derived mast cells, or rat basophilic leukemia cells by either IgE-Ag or calcium ionophore A23187 resulted in inhibition of their proliferation as measured by 3H-thymidine incorporation and MTT (tetrazolium) assays, and in accumulation of the mRNAs of c-fos, c-jun, junB and slightly of junD proto-oncogenes. The involvement of protein kinase C (PKC) in these responses was investigated by using several approaches of enzyme activity regulation. Direct activation of the PKC was achieved by short-term exposure of the cells to the PKC-specific activator phorbol 12-myristate-13-acetate (PMA). Inhibition of PKC activity was obtained by either prolonged treatment of the cells with PMA or by exposure of the cells to the PKC inhibitors H-7 and staurosporine. The results showed the following: (1) Short-term exposure of mast cells to PMA was sufficient to induce inhibition of proliferation. (2) An increase in PKC activity was associated with a decrease in the proliferation of IgE-dinitrophenol (DNP) or calcium ionophore A23187-stimulated cells. (3) A direct correlation was found between the increase in PKC activity and the increase in the level of the mRNAs of the jun proto-oncogenes in cells activated by both stimuli mentioned. (4) While an increase in PKC activity was associated with the upregulation of the level of c-fos mRNA during calcium ionophore A23187 stimulation, it showed the opposite effect on the expression of the mRNA of this proto-oncogene when the cells were triggered by IgE-DNP. Therefore, we concluded that PKC plays various roles in the expression of the mRNA of c-fos in activated mast cells depending on the stimulus involved. In addition, the expression of the mRNA of c-jun and junB proto-onogenes is not coordinately regulated with that of c-fos during immunologic stimulation. This discordancy, which is associated with the increase in PKC activity in mast cells, may play a role in the regulation of the transcription of AP-1-responsive genes, and therefore could be associated with the regulation of proliferation of these cells.


Field transfer decay and interferometry in coupled waveguides with stochastic sources.
Coupled waveguides, such as those used as optical filters, switches, or couplers, are investigated in the context of coherence theory. A stochastic source is introduced to the standard coupled-mode model. Decay in the power transferred between the waveguides as a function of distance is predicted. Application of this result in the form of an interferometric spectrometer is explored.


Crystal Structure of the New Investigational Drug Candidate VT-1598 in Complex with Aspergillus fumigatus Sterol 14α-Demethylase Provides Insights into Its Broad-Spectrum Antifungal Activity.
Within the past few decades, the incidence and complexity of human fungal infections have increased, and therefore, the need for safer and more efficient, broad-spectrum antifungal agents is high. In the study described here, we characterized the new tetrazole-based drug candidate VT-1598 as an inhibitor of sterol 14α-demethylase (CYP51B) from the filamentous fungus Aspergillus fumigatus VT-1598 displayed a high affinity of binding to the enzyme in solution (dissociation constant, 13 ± 1 nM) and in the reconstituted enzymatic reaction was revealed to have an inhibitory potency stronger than the potencies of all other simultaneously tested antifungal drugs, including fluconazole, voriconazole, ketoconazole, and posaconazole. The X-ray structure of the VT-1598/A. fumigatus CYP51 complex was determined and depicts the distinctive binding mode of the inhibitor in the enzyme active site, suggesting the molecular basis of the improved drug potency and broad-spectrum antifungal activity. These data show the formation of an optimized hydrogen bond between the phenoxymethyl oxygen of VT-1598 and the imidazole ring nitrogen of His374, the CYP51 residue that is highly conserved across fungal pathogens and fungus specific. Comparative structural analysis of A. fumigatus CYP51/voriconazole and Candida albicans CYP51/VT-1161 complexes supports the role of H bonding in fungal CYP51/inhibitor complexes and emphasizes the importance of an optimal distance between this interaction and the inhibitor-heme iron interaction. Cellular experiments using two A. fumigatus strains (strains 32820 and 1022) displayed a direct correlation between the effects of the drugs on CYP51B activity and fungal growth inhibition, indicating the noteworthy anti-A. fumigatus potency of VT-1598 and confirming its promise as a broad-spectrum antifungal agent.


Toxicity and efficacy of radioimmunotherapy in carcinoembryonic antigen-producing medullary thyroid cancer xenograft: comparison of iodine 131-labeled F(ab')2 and pretargeted bivalent hapten and evaluation of repeated injections.
This study compared the toxicity and efficacy of 131I-labeled bivalent hapten pretargeted by anti-carcinoembryonic antigen (CEA)/anti-N alpha-(diethylenetriamine-N,N,N',N''-tetraacetic acid-indium(F6-734) bispecific antibody [affinity enhancement system (AES) reagents] with 131I-labeled anti-CEA F(ab')2 (131I-F6) in mice grafted with a human medullary thyroid carcinoma. Repeated injections of AES reagents were also evaluated. Mice bearing TT tumor xenografts were treated with 37, 74, or 92.5 MBq of AES reagents, two injections of 74 MBq of AES reagents 45 days apart, or 37 or 92.5 MBq of 131I-F6. Control groups were treated with nonspecific 131I-labeled F(ab')2, nonspecific AES reagents, nonradiolabeled F6, F6-734 bispecific antibody, and nonradiolabeled bivalent hapten or received no injection. For AES treatments, bispecific antibody was injected 48 h before the hapten. Animal weight, hematological toxicity, tumor volume, and serum thyrocalcitonin were monitored during 5 months. At 92.5 MBq, weight loss was significantly lower after AES than F6 treatment (P = 0.004). The percentages of leukocyte count changes were significantly lower after AES than F6 at 37 and 92.5 MBq (P = 0.01 and 0.04, respectively). The percentage of platelet count changes was significantly lower with AES at the 92.5-MBq dose level (P = 0.04). In the group injected twice with AES reagents, toxicity was not significantly increased after the second treatment. Tumor response was observed in all cases but was significantly longer with repeated treatments of 74 MBq AES reagents than with a single treatment (P = 0.004). Two complete responses were observed with repeated treatments. Changes in thyrocacitonin level paralleled those in tumor volume. These results indicate that pretargeted radioimmunotherapy was at least as efficient as one-step radioimmunotherapy and markedly less toxic. Repeated treatments with AES reagents increased efficacy without increasing toxicity.


Phenotypic characterization of a glucose transporter null mutant in Leishmania mexicana.
Glucose is a major source of energy and carbon in promastigotes of Leishmania mexicana, and its uptake is mediated by three glucose transporters whose genes are encoded within a single cluster. A null mutant in which the glucose transporter gene cluster was deleted by homologous gene replacement was generated previously and shown to grow more slowly than wild type promastigotes but not to be viable as amastigotes in primary tissue culture macrophages or in axenic culture. Further phenotypic characterization demonstrates that the null mutant is unable to import glucose, mannose, fructose, or galactose and that each of the three glucose transporter isoforms, LmGT1, LmGT2, and LmGT3, is capable of transporting each of these hexoses. Complementation of the null mutant with each isoform is able to restore growth in each of the four hexoses to wild type levels. Null mutant promastigotes are reduced in size to about 2/3 the volume of wild type parasites. In addition, the null mutants are significantly more sensitive to oxidative stress than their wild type counterparts. These results underscore the importance of glucose transporters in the parasite life cycle and suggest reasons for their non-viability in the disease-causing amastigote stage.


What caused the epidemic of Pneumocystis pneumonia in European premature infants in the mid-20th century?
An epidemic of interstitial pneumonia principally involving premature infants occurred in Germany and nearby European countries between the 1920s and 1960s. Fatalities were due to Pneumocystis. Because the principal defenses against Pneumocystis are T cells, an acquired T-cell deficiency was postulated. A number of potential causes including malnutrition were considered. All were implausible except for a retrovirus that was benign in adults but virulent in premature infants. Furthermore, we suspect that the virus was imported into Germany from former German African colonies. Premature infants were vulnerable because of the developmental status of their T cells. Given the practices in that part of Europe at that time, the virus was most likely transmitted by contaminated blood transfusions and subsequent contamination of reusable needles and syringes used in injections. Although the epidemic ended 4 decades ago, a search for the postulated retrovirus can be conducted if tissues from affected infants are available.


Coronary artery ectasia presenting with acute inferior wall myocardial infarction in a young adult.
Coronary artery ectasia (CAE), a variant of coronary artery anomalies, is a rare clinical entity. Although atherosclerotic coronary artery disease (CAD) is the most common cause of acute coronary syndrome (ACS), CAE also conveys a relatively high risk for potentially life-threatening cardiac events. We report a case of a 35-year-old male with two-vessel CAE, who presented with acute inferior wall ST election myocardial infarction (STEMI). After initiating medical therapy including bivalirudin, emergent percutaneous coronary intervention (PCI) with stenting of postero-lateral branch of right coronary artery (RCA) was performed. Coronary angiography also showed diffuse ectasia of RCA and left anterior descending artery (LAD). The patient tolerated the procedure well and was discharged on appropriate medical therapy. He was followed-up at one month, with no resulting cardiac events. This case highlights the importance of CAE awareness. CAE has drawn the attention of clinicians because of its clinical implications, as well as its seemingly higher prevalence, due to the abundant use of coronary angiograms and advanced cardiac imaging in the contemporary world. Albeit there has been much progress in the understanding and management of the disease, questions still remain regarding the exact pathophysiology, management guidelines and prognosis of CAE, which are worth further study.


Large-scale movements in European badgers: has the tail of the movement kernel been underestimated?
Characterizing patterns of animal movement is a major aim in population ecology, and yet doing so at an appropriate spatial scale remains a major challenge. Estimating the frequency and distances of movements is of particular importance when species are implicated in the transmission of zoonotic diseases. European badgers (Meles meles) are classically viewed as exhibiting limited dispersal, and yet their movements bring them into conflict with farmers due to their potential to spread bovine tuberculosis in parts of their range. Considerable uncertainty surrounds the movement potential of badgers, and this may be related to the spatial scale of previous empirical studies. We conducted a large-scale mark-recapture study (755 km(2); 2008-2012; 1935 capture events; 963 badgers) to investigate movement patterns in badgers, and undertook a comparative meta-analysis using published data from 15 European populations. The dispersal movement (>1 km) kernel followed an inverse power-law function, with a substantial 'tail' indicating the occurrence of rare long-distance dispersal attempts during the study period. The mean recorded distance from this distribution was 2.6 km, the 95 percentile was 7.3 km and the longest recorded was 22.1 km. Dispersal frequency distributions were significantly different between genders; males dispersed more frequently than females, but females made proportionally more long-distance dispersal attempts than males. We used a subsampling approach to demonstrate that the appropriate minimum spatial scale to characterize badger movements in our study population was 80 km(2), substantially larger than many previous badger studies. Furthermore, the meta-analysis indicated a significant association between maximum movement distance and study area size, while controlling for population density. Maximum long-distance movements were often only recorded by chance beyond the boundaries of study areas. These findings suggest that the tail of the badger movement distribution is currently underestimated. The implications of this for understanding the spatial ecology of badger populations and for the design of disease intervention strategies are potentially significant.


Endoscopic ultrasound, secretin endoscopic pancreatic function test, and histology: correlation in chronic pancreatitis.
Endoscopic ultrasound (EUS) and hormone-stimulated pancreatic function tests are considered useful, and possibly complementary, in the diagnosis of early chronic pancreatitis (CP). Few past studies have compared either methods with a histological gold standard. The aims were to assess correlations of EUS score and endoscopic pancreatic function test (ePFT) results with the degree of histological fibrosis, as well as the sensitivity of each method for detecting fibrosis. This was a retrospective study of patients who underwent EUS, ePFT, or both within 12 months of pancreatic resection or wedge biopsy. EUS scoring was performed using 9 standard criteria, with ≥4 considered abnormal. An ePFT peak bicarbonate concentration <80 mM was considered abnormal. Surgical specimens were reviewed in a blinded manner by an expert pancreatic pathologist and assigned a fibrosis score from 0 to 12. Correlations of the EUS score and ePFT peak bicarbonate with the fibrosis score are reported using the Spearman correlation coefficient. Sensitivity and specificity was calculated for each method against the histological gold standard (fibrosis score ≥2). Twenty-five patients were included. The fibrosis score significantly correlated with the EUS score (r=0.72; 95% confidence interval (CI)=0.43, 0.87; P<0.001) and the ePFT peak bicarbonate (r=-0.57; 95% CI=-0.81, -0.10; P=0.016). EUS had a sensitivity of 84% (95% CI=69, 100) and specificity of 100% (95% CI=40, 100) compared with histology. The ePFT had a sensitivity of 86% (95% CI=67, 100) and specificity of 67% (95% CI=13, 100). When both modalities were combined, the sensitivity increased to 100% (95% CI=63, 100). Both EUS and ePFT are useful tests in the diagnosis of CP. Combining EUS with ePFT may improve the sensitivity for detection of early fibrosis.


Pan-leukocyte monoclonal antibody L3B12. Characterization and application to research and diagnostic problems.
In this report, the authors describe a murine anti-human monoclonal antibody, L3B12, which defines a pan-leukocyte cell surface antigen of approximately 180,000 m.w. Extensive screening against a variety of tissues indicates that L3B12 is sensitive and specific for leukocytes, related cells of bone marrow lineage, and their corresponding neoplasms. Unlike many lymphoid antigens that are not detectable following routine fixation and embedding, those recognized by L3B12 and related antibodies are variably preserved. L3B12 has proven useful in studying the antigen expression of normal leukocytic elements, lymphomas, and related disorders, and in enriching or depleting leukocytes from heterogeneous cell populations. From a diagnostic standpoint, L3B12 staining of tissue sections or cell suspensions is useful for distinguishing large cell lymphomas from undifferentiated carcinomas and in distinguishing lymphomas and leukemias from other small round cell tumors of childhood.


[Molecular mechanism of cardiovascular damage induced by aldosterone].
Although the pro-inflammatory and pro-fibrotic actions of aldosterone on the vasculature have been reported, the effects and molecular mechanisms of aldosterone on endothelial function are yet to be determined. We investigated how aldosterone regulates endothelial nitric oxide synthase (eNOS) function in human umbilical vein endothelial cells (HUVECs). HUVECs were incubated for 16 hrs with 10(-7) mol/l of aldosterone. The concentration of reactive oxygen species (ROS) was estimated by measuring DCF chemiluminescence. Signal transduction was estimated by Western immunoblots. Realtime RT-PCR was performed to measure expression of transcripts of endogenous GTP cyclohydrolase-1 (GCH1) and components of NAD(P)H oxidase. In order to eliminate the possible effect of the glucocorticoid receptor (GR), and to emphasize the role of mineralocorticoid receptor (MR), we used GR siRNA and knocked down GR expression in several experiments. NO output was estimated by intracellular cGMP concentration. ROS production increased significantly in aldosterone-treated HUVEC, but was abolished by pre-treatment with eplerenone. Transcripts of p47(phox) were increased by aldosterone treatment. Vascular endothelial growth factor (VEGF)-induced eNOS Ser 1177 but not Akt Ser 473 phosphorylation levels were reduced significantly by pretreatment with aldosterone. Pretreatment with either eplerenone or okadaic acid restored phosphorylation levels of eNOS Ser 1177 in aldosterone-treated cells, suggesting that protein phosphatase (PP) 2A was upregulated by aldosterone via MR. The decrease in NO output caused by aldosterone pretreatment was reversed significantly by either 5,6,7,8-tetrahydrobiopterin (BH(4)), GCH1 overexpression, or p47(phox) knockdown. These results suggest that aldosterone inhibits eNOS function through bimodal mechanisms of BH(4) deficiency and PP2A activation.


Cumulative Blood Pressure in Early Adulthood and Coronary Artery Calcium and Carotid Intima-Media Thickness in Middle Age Among Adults With Maintained Blood Pressure of <130/80 mm Hg: A Post Hoc Analysis.
To examine the association of blood pressure (BP) levels with coronary artery calcium and carotid intima-media thickness (CIMT) in people with maintained BP below the hypertension range based on current definitions. In this post hoc analysis of the CARDIA (Coronary Artery Risk Development in Young Adults) prospective observational cohort study conducted in 4 US cities, we examined 1233 study participants (mean [SD] age at year 20 examination was 45.3 [3.5] years; 65.4% women). Participants with BP assessments across 20 years and untreated BP of <130/80 mm Hg were included. Multivariable logistic or linear regression models, adjusted for age, sex, race, education, diabetes, body mass index, serum creatinine, smoking, alcohol intake, physical activity, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and triglycerides, were used to examine the associations between cumulative BP measures with coronary artery calcium and CIMT. Higher long-term cumulative systolic BP and pulse pressure across early adulthood were associated with higher CIMT (both P<0.001) but not coronary artery calcium in the multivariable-adjusted model. The associations remained significant even after adjustment for a single BP measurement at year 0 or year 20. The odds ratio (OR) of a maximal CIMT >1.01 mm was ≈50% higher per 1-SD increase in systolic BP (OR, 1.50 [95% CI, 1.19-1.88]) and pulse pressure (OR, 1.46 [95% CI, 1.19-1.79]). Similar findings for CIMT were observed among individuals with a coronary artery calcium score of 0 as well as those with maintained BP of <120/80 mm Hg throughout young adulthood. Long-term cumulative systolic BP and pulse pressure across early adulthood within the nonhypertensive range were associated with adverse midlife alterations in CIMT.


Autoclaved tumor bone for skeletal reconstruction in paediatric patients: a low cost alternative in developing countries.
We reviewed in this series forty patients of pediatric age who underwent resection for malignant tumors of musculoskeletal system followed by biological reconstruction. Our surgical procedure for reconstruction included (1) wide en bloc resection of the tumor; (2) curettage of tumor from the resected bone; (3) autoclaving for 8 minutes (4) bone grafting from the fibula (both vascularized and nonvascularized fibular grafts used); (5) reimplantation of the autoclaved bone into the host bone defect and fixation with plates. Functional evaluation was done using MSTS scoring system. At final followup of at least 18 months (mean 29.2 months), 31 patients had recovered without any complications. Thirty-eight patients successfully achieved a solid bony union between the graft and recipient bone. Three patients had surgical site infection. They were managed with wound debridement and flap coverage of the defect. Local recurrence and nonunion occurred in two patients each. One patient underwent disarticulation at hip due to extensive local disease and one died of metastasis. For patients with non-union, revision procedure with bone graft and compression plates was successfully used. The use of autoclaved tumor grafts provides a limb salvage option that is inexpensive and independent of external resources and is a viable option for musculoskeletal tumor management in developing countries.


Using genomics to understand inter- and intra- outbreak diversity of Pasteurella multocida isolates associated with fowl cholera in meat chickens.
Fowl cholera, caused by Pasteurella multocida, continues to be a challenge in meat-chicken-breeder operations and has emerged as a problem for free-range meat chickens. Here, using whole-genome sequencing (WGS) and phylogenomic analysis, we investigate isolate relatedness during outbreaks of fowl cholera on a free-range meat chicken farm over a 5-year period. Our genomic analysis revealed that while all outbreak isolates were sequence type (ST) 20, they could be separated into two distinct clades (clade 1 and clade 2) consistent with difference in their lipopolysaccharide (LPS) type. The isolates from the earlier outbreaks (clade 1) were carrying LPS type L3 while those from the more recent outbreaks (clade 2) were LPS type L1. Additionally, WGS data indicated high inter- and intra-chicken genetic diversity during a single outbreak. Furthermore, we demonstrate that while a killed autogenous vaccine carrying LPS type L3 had been successful in protecting against challenge from L3 isolates it might have driven the emergence of the closely related clade 2, against which the vaccine was ineffective. The genomic results also revealed a 14 bp deletion in the galactosyltransferase gene gatG in LPS type L3 isolates, which would result in producing a semi-truncated LPS in those isolates. In conclusion, our study clearly demonstrates the advantages of genomic analysis over the conventional PCR-based approaches in providing clear insights in terms of linkage of isolate within and between outbreaks. More importantly, it provides more detailed information than the multiplex PCR on the possible structure of outer LPS, which is very important in the case of strain selection for killed autogenous vaccines.


Progression of liver fibrosis in women infected with hepatitis C: long-term benefit of estrogen exposure.
Female sex is a protective factor for the progression of fibrosis in patients with chronic hepatitis C virus (HCV) infection. Experimental data suggest that estrogens may have an antifibrotic effect. The objective of this study was to evaluate the influence of past pregnancies, oral contraceptives, menopause, and hormone replacement therapy (HRT) on liver fibrosis progression in HCV-infected women. Four hundred seventy-two HCV-infected women received a survey regarding prior pregnancies, menopause, and the use of oral contraceptives and HRT. The impact of these variables on liver fibrosis and its progression were evaluated using multivariate analyses considering all putative confounding factors. Two hundred one women completed the survey (43% response rate), 157 of whom had an estimated date of HCV infection (96 postmenopausal women, 96 women with previous pregnancies, and 105 women with past use of oral contraceptives). Through multivariate analyses, the estimated rate of fibrosis progression was higher in postmenopausal (P < .05) and nulliparous (P = .02) women and was associated with greater histological activity (P < .001). Prior use of oral contraceptives had no significant influence. Among postmenopausal women, the estimated rate of fibrosis progression (+/-SE) was lower in women who received HRT compared with untreated patients (0.099 +/- 0.016 vs. 0.133 +/- 0.006 METAVIR units/yr; P = .02) and was similar to that of premenopausal women (0.093 +/- 0.012 METAVIR units/yr; P value not significant). In conclusion, menopause appears to be associated with accelerated liver fibrosis progression in HCV-infected women, an effect that may be prevented by HRT. Pregnancies may have a beneficial impact on the long-term progression of liver fibrosis.


Up-Regulated Expression of Pro-Apoptotic Long Noncoding RNA lincRNA-p21 with Enhanced Cell Apoptosis in Lupus Nephritis.
Accelerated cell apoptosis with dysregulated long noncoding RNAs is the crucial pathogenesis in lupus nephritis (LN). Pro-apoptotic lincRNA-p21 was studied in LN patients, cell lines with lentivirus-mediated overexpression and CRISPR interference (CRISPRi)-conducted repression, and a mouse model. Clinical samples were from patients and age/sex-matched controls. Expression of lincRNA-p21 and endogenous RNA target miR-181a, were examined in mononuclear and urine cells. Guide RNA sequences targeting lincRNA-p21 were cloned into CRISPRi with dCas9/ Krüppel-associated box (KRAB) domain. LincRNA-p21-silened transfectants were investigated for apoptosis and miR-181a expression. LincRNA-p21-overexpressed cells were evaluated for apoptosis and p53-related down-stream molecules. Balb/C mice were injected with pristane to induce LN and examined for apoptosis and lincRNA-p21. Higher lincRNA-p21 levels were found in LN mononuclear and urine cells, positively correlated with activity. There were lower miR-181a levels in LN mononuclear cells, negatively correlated with activity. Doxorubicin-induced apoptotic cells had up-regulated lincRNA-p21 levels. CRISPRi with dCas9/KARA domain showed efficient repression ability on transcription initiation/elongation. CRISPRi-conducted lincRNA-p21-silenced transfectants displayed reduced apoptosis with up-regulated miR-181a levels, whereas lentivirus-mediated lincRNA-p21-overexpressed cells revealed enhanced apoptosis with up-regulated downstream PUMA/Bax expression. LN mice had glomerular apoptosis with progressive increased lincRNA-p21 levels. Our results demonstrate up-regulated lincRNA-p21 expression in LN, implicating a potential diagnostic marker and therapeutic target.


Endometrial changes with chronic tamoxifen use.
Tamoxifen is a nonsteroidal antiestrogenic drug used successfully to reduce recurrences in all stages of breast carcinoma. Recent reports have emphasized an increased incidence of premalignant and neoplastic endometrial changes that have questioned the risk-benefit ratio of this medication. It appears that the use of tamoxifen increases the incidence of endometrial abnormalities in 1.2 individuals per 1000 women treated with tamoxifen, increasing to 6.3 per 1000 women after 5 years. However, current evidence suggests that the use of tamoxifen as an adjuvant for breast carcinoma overrides the risk of developing endometrial neoplasias. It is not clear which long-term method of surveillance is most cost-effective for these patients. Therefore, individuals should be alerted to the potential risks of tamoxifen use. At present, the screening method of choice should be decided by each individual practitioner, bearing in mind that any vaginal bleeding should be readily investigated. If high-grade hyperplasias of the endometrium or cancer are found, management should be tailored in conjunction with the physician responsible for the woman's breast care.


Predictive role of neutrophil gelatinase-associated lipocalin in early diagnosis of platin-induced renal injury.
Acute kidney injury is an important issue in chemotherapy receiving patients an neutrophil gelatinase-associated lipocalin has been proposed as a novel marker. We here aimed to assess the role of urinary levels for assessment after platin exposure. Patients who had treated with cisplatin or carboplatin or oxaliplatin containg regimens were included in this study. Baseline and postchemotherapy serum urea, creatinine, urine neutrophil gelatinase-associated lipocalin and urine creatinine levels were determined. To avoid the effects of hydration during chemotherapy infusion the urinary neutrophil gelatinase-associated lipocalin/urine creatinine ratio was used to determine acute kidney injury. Of a total of 42 patients receiving platin compounds,14 (33.3%) received cisplatin containing regimens, 14 (33.3%) received carboplatin and 14 (33.3%) oxaliplatin. The median age was 60 (37-76) years. Nineteen of the patients (45.2%) had lung cancer, 12 (28.6%) colorectal cancer and 11 (26.2%) others. The median pre and post chemotherapy urine neutrophil gelatinase-associated lipocalin/urine creatinin ratio was 15.6 ng/mg and 35.8 ng/mg (p=0.041) in the cisplatin group, 32.5 ng/mg and 86.3 ng/mg (p=0.004) in the carboplatin group and 40.9 ng/mg and 62.3 ng/ mg (p=0.243) in the oxaliplatin group. Nephrotoxicity is a serious side effect of chemotherapeutic agentslike cisplatin and carbopaltin, but only to a lower extent oxaliplatin. All platin compounds must be used carefully and urine neutrophil gelatinase-associated lipocalin measurement seems to be promising in detecting acute kidney injury earlier than with creatinine.


Flavour-enhanced cortisol release during gum chewing.
There is some evidence to suggest that chewing gum reduces chronic stress. However, it remains controversial how the taste and odour properties of chewing gum influence stress. The present study was designed to investigate this issue in human subjects. Using an enzyme-linked immunosorbent assay, we tested salivary cortisol concentration, which is thought to be a stress marker, in 96 adults who chewed gum with different combinations of taste and odour. Subjects could discriminate between the types of gum without prior information. Salivary cortisol concentrations were highest and lowest for the subjects who chewed the most flavourful gum and the least flavourful gum, respectively. These findings suggest that the salivary cortisol level during gum chewing is not a marker of negative emotions (i.e., stressful conditions) as traditionally considered but, rather, an index of positive emotions that can facilitate biological responses to overcome stressful conditions.


Robot-assisted nephroureterectomy for upper tract urothelial carcinoma: results from three high-volume robotic surgery institutions.
Robot-Assisted NephroUreterectomy (RANU) represents a minimally invasive alternative to open NephroUreterectomy (NU) for management of Upper Tract Urothelial Carcinoma (UTUC) but its oncologic safety is still controversial. The objective of this study was to investigate the peri-operative, pathologic and oncologic outcomes of RANU for UTUC. From 2008 to 2017, 78 patients diagnosed with UTUC and elected for RANU at 3 high-volume robotic surgery centres were retrospectively assessed. Surgery was performed using da Vinci Si® and Xi® systems. RANU was done adhering to oncological principles as in open surgery. The outcomes of the study were: (1) peri-operative morbidity, namely intra- and post-operative complications, blood loss, length of hospital stay and operative time; (2) oncologic outcomes, namely overall survival (OS) and recurrence-free survival (RFS). Peri-operative overall complication rate was 24.4% and high-grade complication rate was 2.6%. Median blood loss, length of hospital stay and operative time were 124 ml, 4 days and 167 min. Lymphadenectomy was performed in 31 (41%) patients. Lymph-node involvement was present in 9 (29%) patients. At median follow-up of 15 months, 2- and 4-year OS were 79% and 66%, respectively, and RFS was 63% and 53%. Peritoneal dissemination was recorded in 1 (1.3%) patient with pT4N2R1 UTUC. Our study is limited by the relatively small cohort of patients and its retrospective character. RANU as minimally invasive treatment for patients with UTUC is safe and feasible. Post-operative morbidity is low and major complications are rare. Oncologic outcomes are acceptable and no evidence of increased risk of peritoneal dissemination is recorded. Long-term data are needed. RANU should be regarded as an alternative to open surgery for UTUC that can offer good peri-operative and oncologic results.


Increased level of p21 in human ovarian tumors is associated with increased expression of cdk2, cyclin A and PCNA.
We have demonstrated over-expression of the cyclin-dependent kinase inhibitor p21 in various ovarian-cancer cell lines as well as in ovarian-tumor biopsies. This increase in p21 expression relative to that observed in normal ovarian epithelial cells is unrelated to proliferation index. In the present study, we found that p21 is functional, since the protein extracted from IGROVI cells is still able to inhibit cdk2-kinase activity. We then investigated how IGROVI cells overcome the growth-inhibitory function of p21. Immunofluorescence assays and subcellular fractionation showed that p21 is located in cytoplasm and nucleus both in normal and in tumoral cells. Compared with normal ovarian epithelial cells in culture, the increase in level of p21 in IGROVI cells was found to be associated with increased expression of cdk2, cyclin-A and PCNA proteins. In IGROVI cells, p21 is associated with inactive cdk2/cyclin-A complex, indicating that it acts as an inhibitory factor rather than an assembly factor. Over-expression of cdk2 and of cyclin A observed in IGROVI cells allows them to escape to p21-inhibitory activity. The fact that cells from ovarian-tumor biopsies exhibited a concomitant increase in p21 and in its partners cdk2 and PCNA suggest that ovarian-tumor cells can tolerate high levels of functional p21 via over-expression of other cell-cycle-regulatory proteins.


Some Gammaproteobacteria are enriched within CD14+ macrophages from intestinal lamina propria of Crohn's disease patients versus mucus.
Crohn's disease causes chronic inflammation in the gastrointestinal tract and its pathogenesis remains unclear. In the intestine of Crohn's disease patients, CD14+CD11+CD163low macrophages contribute to inflammation through the induction of Th17 cells and production of inflammatory cytokines; the CD14+CD11c+163high fraction is anti-inflammatory through the production of IL-10 in normal cases. In this report, the 16S rRNA gene amplicon sequencing method was used to identify bacteria that are specifically present in intestinal CD14+CD11c+ macrophages of Crohn's disease patients. Bacteria present in intestinal CD14+CD11c+ macrophages and mucus of Crohn's disease patients were separated into different clusters in principal coordinates analysis. There was a statistically significant increase in the relative composition of CD14+CD11c+ macrophages from mucus in two phyla (Proteobacteria [p = 0.01] and Actinobacteria [p = 0.02]) and two families (Moraxellaceae [p < 0.001] and Pseudomonadaceae [p = 0.01]). In addition, OTU-1: Acinetobacter and OTU-8: Pseudomonadaceae tended to concentrate in the CD14+CD11c+CD163low subset, whereas OTU-10: Proteus, OTU-15: Collinsella tended to concentrate more in the CD14+CD11c+CD163high subset than the other subset and mucus.


Monocular blindness during therapy for cerebral neurocysticercosis.
Neurocysticercosis is endemic in India, cerebral and ocular manifestations being common. A 32 yr old man on treatment with Albendazole for cerebral neurocysticercosis for 10 days presented with 3 days of painful uniocular blindness. He had only light perception in the left eye, left pupil was non-reactive to light and left disc was edematous. B-scan of eye revealed retinal detachment due to sub retinal cyst and CT brain showed multiple parenchymal cysticerci. The natural history of ocular neurocysticercosis or enhanced sub-retinal inflammation due to Albendazole therapy could have resulted in the retinal detachment in this case.


Emerging Parasitic Protists: The Case of Perkinsea.
The last century has witnessed an increasing rate of new disease emergence across the world leading to permanent loss of biodiversity. Perkinsea is a microeukaryotic parasitic phylum composed of four main lineages of parasitic protists with broad host ranges. Some of them represent major ecological and economical threats because of their geographically invasive ability and pathogenicity (leading to mortality events). In marine environments, three lineages are currently described, the Parviluciferaceae, the Perkinsidae, and the Xcellidae, infecting, respectively, dinoflagellates, mollusks, and fish. In contrast, only one lineage is officially described in freshwater environments: the severe Perkinsea infectious agent infecting frog tadpoles. The advent of high-throughput sequencing methods, mainly based on 18S rRNA assays, showed that Perkinsea is far more diverse than the previously four described lineages especially in freshwater environments. Indeed, some lineages could be parasites of green microalgae, but a formal nature of the interaction needs to be explored. Hence, to date, most of the newly described aquatic clusters are only defined by their environmental sequences and are still not (yet) associated with any host. The unveiling of this microbial black box presents a multitude of research challenges to understand their ecological roles and ultimately to prevent their most negative impacts. This review summarizes the biological and ecological traits of Perkinsea-their diversity, life cycle, host preferences, pathogenicity, and highlights their diversity and ubiquity in association with a wide range of hosts.


The Bilateral Carotid Artery Stenosis (BCAS) Model of Vascular Dementia.
Vascular dementia is the second most common form of dementia after Alzheimer's disease. Chronic cerebral hypoperfusion is a key contributor to the development of vascular dementia. In this chapter, we describe the surgical procedures used for bilateral carotid artery stenosis (BCAS) surgery to induce chronic cerebral hypoperfusion. Mice that undergo BCAS surgery develop the hallmarks of vascular dementia including white matter lesions, neuroinflammation, and cognitive impairment. This technique may be used for studies of chronic cerebral hypoperfusion and vascular dementia in mice.


Individual Differences in the Contribution of Maternal Soothing to Infant Distress Reduction.
This study investigates individual differences in the contribution of specific maternal regulatory behaviors to the mother-infant dyad's regulation of infant distress response. Additionally, we examined the stability of infants' stress responses and the stability of specific maternal soothing behaviors. The sample included 128 mother-infant dyads that were observed during an inoculation at 2 and 6 months. The average intensity of infant cry response showed modest stability across age only before controlling for the infant's general state of irritability, and the duration of crying was not stable. Of the 8 specific maternal regulatory behaviors studied, affection, touching, and vocalizing showed the strongest stability across infant age. Finally, an index of the contingency between maternal soothing and infant cry reduction at 2 months predicted shorter cry duration but not cry intensity at 6 months. The results of this study indicate that infants whose mothers showed a greater contribution to reducing their distress at 2 months showed a shorter duration of crying 4 months later. This suggests a possible longitudinal influence of maternal regulation on infants' distress responses.


A framework for gene mapping in wheat demonstrated using the Yr7 yellow rust resistance gene.
We used three approaches to map the yellow rust resistance gene Yr7 and identify associated SNPs in wheat. First, we used a traditional QTL mapping approach using a double haploid (DH) population and mapped Yr7 to a low-recombination region of chromosome 2B. To fine map the QTL, we then used an association mapping panel. Both populations were SNP array genotyped allowing alignment of QTL and genome-wide association scans based on common segregating SNPs. Analysis of the association panel spanning the QTL interval, narrowed the interval down to a single haplotype block. Finally, we used mapping-by-sequencing of resistant and susceptible DH bulks to identify a candidate gene in the interval showing high homology to a previously suggested Yr7 candidate and to populate the Yr7 interval with a higher density of polymorphisms. We highlight the power of combining mapping-by-sequencing, delivering a complete list of gene-based segregating polymorphisms in the interval with the high recombination, low LD precision of the association mapping panel. Our mapping-by-sequencing methodology is applicable to any trait and our results validate the approach in wheat, where with a near complete reference genome sequence, we are able to define a small interval containing the causative gene.


Cardiac Safety Research Consortium: can the thorough QT/QTc study be replaced by early QT assessment in routine clinical pharmacology studies? Scientific update and a research proposal for a path forward.
The International Conference on Harmonization E14 guidance for the clinical evaluation of QT/QTc interval prolongation requires almost all new drugs to undergo a dedicated clinical study, primarily in healthy volunteers, the so-called TQT study. Since 2005, when the E14 guidance was implemented in United States and Europe, close to 400 TQT studies have been conducted. In February 2012, the Cardiac Safety Research Consortium held a think tank meeting at Food and Drug Administration's White Oak campus to discuss whether "QT assessment" can be performed as part of routine phase 1 studies. Based on these discussions, a group of experts convened to discuss how to improve the confidence in QT data from early clinical studies, for example, the First-Time-in-Human trial, through collection of serial electrocardiograms and pharmacokinetic samples and the use of exposure response analysis. Recommendations are given on how to design such "early electrocardiogram assessment," and the limitation of not having a pharmacologic-positive control in these studies is discussed. A research path is identified toward collecting evidence to replace or provide an alternative to the dedicated TQT study.


Providing medical evaluations for possible child maltreatment to children with special health care needs.
Children with special health care needs are known to be at increased risk of all forms of child maltreatment when compared to children without such needs. We describe a health care team's experience providing medical evaluations for suspected child maltreatment to children with special health care needs. Consecutive cases seen as outpatients in the Abuse Referral Clinic for Children with Disabilities were abstracted and analyzed. Mail and telephone follow-up contact was attempted after the medical evaluation to determine adherence with treatment recommendations. A subsample of cases for which complete financial information was available was reviewed to determine a reimbursement rate. During the study, 49 children received complete outpatient evaluations. Ages ranged from 3 to 16 years old, and 54% were males. Special needs spanned a wide range of physical, developmental/cognitive and behavioral conditions. The largest number of referrals came from child protective services (42%) followed by referrals from physicians (27%). After the team's comprehensive evaluation, 18% of the children were found to have a history or physical examination that was diagnostic for child maltreatment, 13% were thought to be at high risk, 25% were thought to be at low risk and 44% were thought to have non-abusive etiologies. The collection rate was 14% for an average reimbursement of $38 per case. Only 29 caregivers could be found at follow-up and 22 remembered the recommendations made by the team. Of the 25 cases that were referred for outpatient mental health counseling, 12 (48%) complied. Children with a wide range of special health care needs were evaluated in an outpatient special health care needs clinic that offered comprehensive medical evaluations for possible child maltreatment. Medical evaluation services for this group of children were poorly reimbursed. Mental health services were frequently recommended but often not accessed. Child maltreatment teams seeking to serve children with special health care needs will need to plan for service delivery to a potentially diverse group of children and families who may experience difficulty in carrying through on the team's treatment recommendations.


Minimally invasive surgery in veterinary practice: a 2010 survey of diplomates and residents of the American College of Veterinary Surgeons.
To report the current state of minimally invasive surgery (MIS) in veterinary surgical practice in 2010. Electronic questionnaire. Diplomates and residents of the American College of Veterinary Surgeons (ACVS). A survey (38 questions for Diplomates, 23 questions for residents) was sent electronically to 1216 Diplomates and 300 residents. Questions were organized into 5 categories to investigate: (1) caseload and distribution of MIS cases; (2) MIS training; (3) MIS benefits, morbidity, limitations and motivating factors; (4) ACVS role; and (5) demographics of the study population. Eighty-six percent of small animal (SA) Diplomates, 99% of large animal (LA) Diplomates, and 98% of residents had performed MIS. Median LA caseload (30 cases/year; range, 1-600) was significantly higher than SA caseload (20 cases/year; range, 1-350). Descending order of case distribution was: arthroscopy > laparoscopy > endoscopic upper airway > thoracoscopy. Sixty percent of Diplomates and 98% of residents received MIS training during their residency. Residents' perspective of MIS training proficiency was positively correlated to caseload. Ninety-five percent of all respondents felt postoperative morbidity was less with MIS, and were motivated by patient benefits, maintaining a high standard of care, and personal interests. Fifty-eight percent of Diplomates and 89% of residents felt ACVS should be involved in developing MIS training. MIS is widely used by ACVS Diplomates and residents in clinical practice; however, important differences exist between SA and LA surgeons and practice types. MIS training in partnership with the ACVS is needed for continued development in veterinary surgery.


Diagnostic accuracy of power Doppler ultrasound in patients with chronic tennis elbow.
To determine the diagnostic accuracy of power Doppler and grey scale ultrasonography, assess the relationship between severity measures and neovascularity, and describe the intra-tendon distribution of neovascularity in chronic tennis elbow. Between group cross sectional study. Sports medicine clinic and radiology centre. 32 affected elbows (median and range of duration: 10; 3-120 months) and 18 unaffected contralateral elbows in 25 patients (mean age 50 years) with lateral elbow pain, and 38 unaffected elbows in 19 asymptomatic participants (mean age 45 years) underwent a clinical examination (reference standard test) and grey scale and power Doppler ultrasonographic examination. Ultrasound examination with power Doppler identified neovascularity and grey scale ultrasound changes (lateral epicondyle bony spurring or irregularity, maximum anterior-posterior thickness, and echo characteristics) of the common extensor tendon. Power Doppler had a strong positive likelihood ratio of 45.39, whereas a combined null finding in power Doppler and grey scale ultrasonography resulted in a robust negative likelihood ratio of 0.05. Grey scale changes were generally not as diagnostically accurate. Common extensor tendon neovascularity was equally distributed between the superficial and deep part of the tendon, and clinical severity measures did not correlate with neovascularity scores. Neovascularity identified with power Doppler ultrasonography when compared to grey scale changes (alone or in combination with Doppler) was diagnostically superior in identifying chronic tennis elbow. The lack of both neovascularity and grey scale changes on ultrasound examination also substantially increase the probability that the condition is not present and should prompt the clinician to consider other causes for lateral elbow pain.


Authenticity and the psychology of choice in the analyst.
There is a growing appreciation of an ethic of authenticity in analytic technique, a trend related to a recognition of the engagement of the spontaneous, unconscious dimension of the analyst's mind in the clinical situation. This liberatory trend in our theory of technique can and should be elaborated to take account of situations in which the analyst deliberately and strategically attempts to influence the patient for both analytic and therapeutic purposes. There is a complex, dialectical relationship between intentionally planning to provide mutative relational experiences for a patient and the irreducible emotional responsivity that marks every analytic encounter. It is suggested that the dangers of the patient's compliance with and idealization of the analyst usually associated with the analyst's deliberate enacting of attitudes presumed to be mutative are not necessarily inevitable.


Optimizing levothyroxine treatment for subclinical hypothyroidism during pregnancy.
The onset of pregnancy places additional stress of the thyroid gland, which must produce additional thyroid hormones to support the developing foetus. Hypothyroidism, including subclinical hypothyroidism (SCH), may appear de novo at this time, or existing thyroid disease may become more severe. Accordingly, SCH is a relatively common complication of up to about 3% of pregnancies, with higher rates in some areas. There is strong evidence from systematic reviews and meta-analyses that uncontrolled SCH is associated with an increased risk of adverse pregnancy outcomes, including miscarriage, preeclampsia, and gestational diabetes. The evidence base also suggests that treatment with levothyroxine (LT4), optimized to control thyrotropin (TSH) to within its pregnancy-specific reference ranges reduces these risks. Current management guidelines provide a clear framework of intervention with LT4 in pregnant women with SCH, especially where TSH is high or where thyroperoxidase autoantibodies are present. Sub-optimal adherence to LT4 is common: it is important that patients take their LT4 correctly and that treating physicians and/or healthcare professionals manage these patients according to the latest management guidelines. The titration of LT4 is likely to occur within a range of LT4 daily doses between 25 µg and 75 µg for the majority of this population. LT4 is a narrow therapeutic index drug and small variations in dosage may produce a clinically significant change in thyroid status. Newer formulations of LT4, engineered to provide more precise and consistent dosing, and with a broad range of tablet strengths, may facilitate the precise titration of the LT4 dose for these patients.


Correlation of anthropometric parameters with semitendinosus tendon length in anterior cruciate ligament injured patients.
Anterior cruciate ligament reconstruction (ACLR) surgery is now a common procedure performed following a tear of the anterior cruciate ligament. The length of the harvested hamstring tendon is critical to the outcome of the surgery as it may influence the final length and thickness of the graft. The goal of this study was to attempt to establish a relation- ship between the height, weight and body mass index and the harvested length of the semitendinosus tendon of patients having ACLR surgery. This was a retrospective study. The weight, height, and body mass index (BMI) of patients for primary anterior cruciate ligament reconstruction were noted. The average length of the tendon was noted. The correla- tion between each of these anthropometric parameters and the tendon length was estimated. The range of values for the semitendinosus length, height and weight in males and females respectively were 24-39cm (31.26 +/- 2.93) and 26-35cm (29.26 +/- 2.08); 1.6-1.96 m and 1.65-1.8m; and 52-110kg and 60-106kg. Only the height revealed a moderate correlation with the length of the harvested tendon among the male patients. Linear regression analyses yielded the equation Sem- itendinosus tendon length = 23.25xheight-10.28 at a p-value of 0.002. Patient height could be predictive of the length of the harvested semitendinosus tendon.


Impaired fixation to eyes during facial emotion labelling in children with bipolar disorder or severe mood dysregulation.
Children with bipolar disorder (BD) or severe mood dysregulation (SMD) show behavioural and neural deficits during facial emotion processing. In those with other psychiatric disorders, such deficits have been associated with reduced attention to eye regions while looking at faces. We examined gaze fixation patterns during a facial emotion labelling task among children with pediatric BD and SMD and among healthy controls. Participants viewed facial expressions with varying emotions (anger, fear, sadness, happiness, neutral) and emotional levels (60%, 80%, 100%) and labelled emotional expressions. Our study included 22 children with BD, 28 with SMD and 22 controls. Across all facial emotions, children with BD and SMD made more labelling errors than controls. Compared with controls, children with BD spent less time looking at eyes and made fewer eye fixations across emotional expressions. Gaze patterns in children with SMD tended to fall between those of children with BD and controls, although they did not differ significantly from either of these groups on most measures. Decreased fixations to eyes correlated with lower labelling accuracy in children with BD, but not in those with SMD or in controls. Most children with BD were medicated, which precluded our ability to evaluate medication effects on gaze patterns. Facial emotion labelling deficits in children with BD are associated with impaired attention to eyes. Future research should examine whether impaired attention to eyes is associated with neural dysfunction. Eye gaze deficits in children with BD during facial emotion labelling may also have treatment implications. Finally, children with SMD exhibited decreased attention to eyes to a lesser extent than those with BD, and these equivocal findings are worthy of further study.


Clinical features of unilateral moyamoya disease.
Moyamoya disease is characterized by progressive occlusion of the internal carotid artery or its terminal branches, associated with formation of extensive collateral vessels (moyamoya vessels) at the base of the brain. Whether unilateral moyamoya disease, confirmed by typical angiographic evidence of moyamoya disease unilaterally and normal or equivocal findings contralaterally, is an early form of definite (bilateral) moyamoya disease remains controversial. The present study investigated the incidence and clinical features of unilateral moyamoya disease in a series of patients treated for moyamoya disease. Fifty-two patients were diagnosed with definite moyamoya disease and nine patients with unilateral moyamoya disease. Sex, age, signs at onset, neuroimaging findings, treatment, course of the disease, and family history of unilateral moyamoya disease were reviewed. Among the nine patients with unilateral moyamoya disease, there were twice as many females as males, and mean age at onset was 39.0 years. The clinical presentation was ischemic in three patients, bleeding in one, and asymptomatic in five. Two had familial moyamoya disease. Progression to bilateral lesions is known to occur in pediatric patients and patients with stenotic changes of the contralateral internal carotid artery bifurcation. Some unilateral cases are caused by the same genetic defects as definite cases, and others seem to be an unusual form of stenoocclusive process of cerebral arteries. Surgical treatment on the symptomatic side followed by close observation for bilateral involvement is recommended.


Endothelial matrix assembly during capillary morphogenesis: insights from chimeric TagRFP-fibronectin matrix.
Biologically relevant, three-dimensional extracellular matrix is an essential component of in vitro vasculogenesis models. WI-38 fibroblasts assemble a 3D matrix that induces endothelial tubulogenesis, but this model is challenged by fibroblast senescence and the inability to distinguish endothelial cell-derived matrix from matrix made by WI-38 fibroblasts. Matrices produced by hTERT-immortalized WI-38 recapitulated those produced by wild type fibroblasts. ECM fibrils were heavily populated by tenascin-C, fibronectin, and type VI collagen. Nearly half of the total type I collagen, but only a small fraction of the type IV collagen, were incorporated into ECM. Stable hTERT-WI-38 transfectants expressing TagRFP-fibronectin incorporated TagRFP into ~90% of the fibronectin in 3D matrices. TagRFP-fibronectin colocalized with tenascin-C and with type I collagen in a pattern that was similar to that seen in matrices from wild type WI-38. Human Umbilical Vein Endothelial Cells (HUVEC) formed 3D adhesions and tubes on WI38-hTERT-TagRFP-FN-derived matrices, and the TagRFP-fibronectin component of this new 3D human fibroblast matrix model facilitated the demonstration of concentrated membrane type 1 metalloprotease and new HUVEC FN and collagen type IV fibrils during EC tubulogenesis. These findings indicate that WI-38-hTERT- and WI-38-hTERT-TagRFP-FN-derived matrices provide platforms for the definition of new matrix assembly and remodeling events during vasculogenesis.


Interleukin-driven insulin-like growth factor promotes prostatic inflammatory hyperplasia.
Prostatic inflammation is of considerable importance to urologic research because of its association with benign prostatic hyperplasia and prostate cancer. However, the mechanisms by which inflammation leads to proliferation and growth remain obscure. Here, we show that insulin-like growth factors (IGFs), previously known as critical developmental growth factors during prostate organogenesis, are induced by inflammation as part of the proliferative recovery to inflammation. Using genetic models and in vivo IGF receptor blockade, we demonstrate that the hyperplastic response to inflammation depends on interleukin-1-driven IGF signaling. We show that human prostatic hyperplasia is associated with IGF pathway activation specifically localized to foci of inflammation. This demonstrates that mechanisms of inflammation-induced epithelial proliferation and hyperplasia involve the induction of developmental growth factors, further establishing a link between inflammatory and developmental signals and providing a mechanistic basis for the management of proliferative diseases by IGF pathway modulation.


Excited state dynamics of the isolated green fluorescent protein chromophore anion following UV excitation.
A combined frequency-, angle-, and time-resolved photoelectron spectroscopy study is used to unravel the excited state dynamics following UV excitation of the isolated anionic chromophore of the green fluorescent protein (GFP). The optically bright S3 state, which is populated for hv > 3.7 eV, is shown to decay predominantly by internal conversion to the S2 state that in turn autodetaches to the neutral ground state. For hv > 4.1 eV, a new and favorable autodetachment channel from the S2 state becomes available, which leads to the formation of the neutral in an excited state. The results indicate that the UV excited state dynamics of the GFP chromophore involve a number of strongly coupled excited states.


Participation differences by age and depression 5 years after moderate-to-severe traumatic brain injury.
Participation restrictions, including restrictions in employment, recreational activities, and social interactions, and depression are common after traumatic brain injury (TBI) and can profoundly affect individuals. Participation and depression demonstrate complex relationships with each other and over time as individuals age. This study (1) identified differences in participation between different age groups; (2) determined if participation differed between those with and without clinically significant depressive symptoms within each age group; and (3) determined the effect of the interaction between age groups and the presence or absence of clinically significant depressive symptoms on participation in community-dwelling adults with a moderate-to-severe TBI. Results indicate that, among community-dwelling adults 5 years post-TBI, there are significant differences in participation between age groups across the lifespan, with younger adults generally having higher levels of participation. Individuals with clinically significant depressive symptoms participate less than individuals without it within the same age group, except for adults over 65 years-old. For the productivity domain, age interacted with depressive symptoms, such that the presence of clinically significant depressive symptoms was associated with a larger difference in productivity in early-to-middle adulthood. Based on these findings, depression should be considered when providing interventions for participation and vice versa.


Postgestational lymphotoxin/lymphotoxin beta receptor interactions are essential for the presence of intestinal B lymphocytes.
Lymphotoxin (LT), a cytokine belonging to the TNF family, has established roles in the formation of secondary lymphoid structures and in the compartmentalization of T and B lymphocyte areas of the spleen. In this study, we examine the role of LT in directing the composition of intestinal lymphocytes. We report that mice deficient in LT have a normal composition of intestinal lamina propria (LP) T lymphocytes, and an absence of intestinal LP B lymphocytes. We further refine this observation to demonstrate that the interaction of LT with the LTbetaR is essential for the presence LP B lymphocytes. The LT/LTbetaR-dependent events relevant for the presence of LP B lymphocytes occur after birth, do not require the presence of Peyer's patches, lymph nodes, or the spleen; and therefore, are distinct and independent from the previously identified roles of LT/LTbetaR. The LT-dependent signal relevant for the presence of LP B lymphocytes is optimally supplied by a LT-sufficient B lymphocyte, and requires a LTbetaR-sufficient radio-resistant, non-bone marrow-derived cell. Based upon the severity of the deficit of LP B lymphocytes we observed, these novel LT/LTbetaR-dependent events are of primary importance in directing the entry and residence of LP B lymphocytes.


Efficacy of Carraguard-based microbicides in vivo despite variable in vitro activity.
Anti-HIV microbicides are being investigated in clinical trials and understanding how promising strategies work, coincident with demonstrating efficacy in vivo, is central to advancing new generation microbicides. We evaluated Carraguard and a new generation Carraguard-based formulation containing the non-nucleoside reverse transcriptase inhibitor (NNRTI) MIV-150 (PC-817). Since dendritic cells (DCs) are believed to be important in HIV transmission, the formulations were tested for the ability to limit DC-driven infection in vitro versus vaginal infection of macaques with RT-SHIV (SIVmac239 bearing HIV reverse transcriptase). Carraguard showed limited activity against cell-free and mature DC-driven RT-SHIV infections and, surprisingly, low doses of Carraguard enhanced infection. However, nanomolar amounts of MIV-150 overcame enhancement and blocked DC-transmitted infection. In contrast, Carraguard impeded infection of immature DCs coincident with DC maturation. Despite this variable activity in vitro, Carraguard and PC-817 prevented vaginal transmission of RT-SHIV when applied 30 min prior to challenge. PC-817 appeared no more effective than Carraguard in vivo, due to the limited activity of a single dose of MIV-150 and the dominant barrier effect of Carraguard. However, 3 doses of MIV-150 in placebo gel at and around challenge limited vaginal infection, demonstrating the potential activity of a topically applied NNRTI. These data demonstrate discordant observations when comparing in vitro and in vivo efficacy of Carraguard-based microbicides, highlighting the difficulties in testing putative anti-viral strategies in vitro to predict in vivo activity. This work also underscores the potential of Carraguard-based formulations for the delivery of anti-viral drugs to prevent vaginal HIV infection.


Epidemiology and Recognition of Binge-Eating Disorder in Psychiatry and Primary Care.
Substantial unmet needs exist regarding the awareness, diagnosis, and treatment of binge-eating disorder (BED). Affecting both men and women and appearing in all ethnic groups, BED is the most prevalent of all the eating disorders in the United States and worldwide. Left untreated, BED causes significant impairment, reduced quality of life, and decreased productivity. Many patients are unaware of the disorder and present for treatment of weight-related issues or comorbid medical and psychiatric conditions. Communication barriers, such as the reluctance of patients to volunteer information about their eating habits and of clinicians to ask potentially sensitive questions, may be overcome with the use of diagnostic criteria along with appropriate assessment questions and screening tools. Early recognition and accurate diagnosis may help mitigate the long-term impact of BED.


Molecular interactions of STAC proteins with skeletal muscle dihydropyridine receptor and excitation-contraction coupling.
Excitation-contraction coupling (ECC) is the physiological process in which an electrical signal originating from the central nervous system is converted into muscle contraction. In skeletal muscle tissue, the key step in the molecular mechanism of ECC initiated by the muscle action potential is the cooperation between two Ca2+ channels, dihydropyridine receptor (DHPR; voltage-dependent L-type calcium channel) and ryanodine receptor 1 (RyR1). These two channels were originally postulated to communicate with each other via direct mechanical interactions; however, the molecular details of this cooperation have remained ambiguous. Recently, it has been proposed that one or more supporting proteins are in fact required for communication of DHPR with RyR1 during the ECC process. One such protein that is increasingly believed to play a role in this interaction is the SH3 and cysteine-rich domain-containing protein 3 (STAC3), which has been proposed to bind a cytosolic portion of the DHPR α1S subunit known as the II-III loop. In this work, we present direct evidence for an interaction between a small peptide sequence of the II-III loop and several residues within the SH3 domains of STAC3 as well as the neuronal isoform STAC2. Differences in this interaction between STAC3 and STAC2 suggest that STAC3 possesses distinct biophysical features that are potentially important for its physiological interactions with the II-III loop. Therefore, this work demonstrates an isoform-specific interaction between STAC3 and the II-III loop of DHPR and provides novel insights into a putative molecular mechanism behind this association in the skeletal muscle ECC process.


Emerging delivery approaches for targeted pulmonary fibrosis treatment.
Pulmonary fibrosis (PF) is a progressive, and life-threatening interstitial lung disease which causes scarring in the lung parenchyma and thereby affects architecture and functioning of lung. It is an irreversible damage to lung functioning which is related to epithelial cell injury, immense accumulation of immune cells and inflammatory cytokines, and irregular recruitment of extracellular matrix. The inflammatory cytokines trigger the differentiation of fibroblasts into activated fibroblasts, also known as myofibroblasts, which further increase the production and deposition of collagen at the injury sites in the lung. Despite the significant morbidity and mortality associated with PF, there is no available treatment that efficiently and effectively treats the disease by reversing their underlying pathologies. In recent years, many therapeutic regimens, for instance, rho kinase inhibitors, Smad signaling pathway inhibitors, p38, BCL-xL/ BCL-2 and JNK pathway inhibitors, have been found to be potent and effective in treating PF, in preclinical stages. However, due to non-selectivity and non-specificity, the therapeutic molecules also result in toxicity mediated severe side effects. Hence, this review demonstrates recent advances on PF pathology, mechanism and targets related to PF, development of various drug delivery systems based on small molecules, RNAs, oligonucleotides, peptides, antibodies, exosomes, and stem cells for the treatment of PF and the progress of various therapeutic treatments in clinical trials to advance PF treatment.


Reduced mammalian target of rapamycin activity facilitates mitochondrial retrograde signaling and increases life span in normal human fibroblasts.
Coordinated expression of mitochondrial and nuclear genes is required to maintain proper mitochondrial function. However, the precise mechanisms that ensure this coordination are not well defined. We find that signaling from mitochondria to the nucleus is influenced by mammalian target of rapamycin (mTOR) activity via changes in autophagy and p62/SQSTM1 turnover. Reducing mTOR activity increases autophagic flux, enhances mitochondrial membrane potential, reduces reactive oxygen species within the cell, and increases replicative life span. These effects appear to be mediated in part by an interaction between p62/SQSTM1 and Keap1. This interaction allows nuclear accumulation of the nuclear factor erythroid 2-like 2 (NFE2L2, also known as nuclear factor related factor 2 or NRF2), increased expression of the nuclear respiratory factor 1 (NRF1), and increased expression of nuclear-encoded mitochondrial genes, such as the mitochondrial transcription factor A, and mitochondrial-encoded genes involved in oxidative phosphorylation. These findings reveal a portion of the intracellular signaling network that couples mitochondrial turnover with mitochondrial renewal to maintain homeostasis within the cell and suggest mechanisms whereby a reduction in mTOR activity may enhance longevity.


Isolated splenic metastasis from renal cell carcinoma: case report and review.
This report presents the case of a 70-year-old woman with a previous history of a left nephrectomy for renal cell carcinoma (RCC), who developed general malaise and fatigue. Abdominal computed tomography demonstrated an enhancing 6 × 7 cm necrotic lesion in the lower pole of the spleen suggestive of a metastasis. Given the highly suspicious nature of the lesion we proceeded to splenectomy. The tumour did not breach the splenic capsule, and there was no local diaphragmatic involvement. The mass was concluded to be a true metastasis of the original RCC rather than local recurrence of the disease. The causes of isolated solid splenic lesions are wide and varied, however a past or present history of malignancy should lead to a high index of suspicion for a splenic metastasis. We report an extremely unusual case of spread from a RCC.


COVID-19? Let me see your hands.
During the first wave of the COVID�19 pandemic, a patient with anti-synthetase syndrome (ASS) was misdiagnosed as having bilateral severe acute respiratory syndrome coronavirus 2 pneumonia on admission. A comprehensive clinical evaluation would have led to the correct diagnosis earlier, as he had some data consistent with ASS on both physical examination and laboratory tests that were initially overlooked. In addition, a malignant lesion in the colon was found on screening for underlying malignancy. In this context, ASS has been considered a low-risk subgroup for cancer among idiopathic inflammatory myopathies. However, this should be interpreted cautiously and should not lead to neglect of adequate cancer screening adjusted for age, sex and other potential risk factors.


Acceptability, compliance, and adverse reactions when isoniazid, rifampin, and pyrazinamide are given as a combined formulation or separately during three-times-weekly antituberculosis chemotherapy.
In Hong Kong, 627 adult Chinese patients with newly diagnosed pulmonary tuberculosis received, for the first 2 months of chemotherapy, streptomycin together with isoniazid, rifampin, and pyrazinamide allocated at random to be given either as a combined formulation or separately. Each tablet of the combined formulation, which was designed for intermittent use, contained 125 mg of isoniazid, 100 mg of rifampin, and 375 mg of pyrazinamide. Patients weighing 42 kg or less received 5 tablets per dose, 43 to 57 kg, 6 tablets per dose, and 58 kg or more, 7 tablets per dose. The dosage of each drug was very similar whether it was given combined or separately. During the 2 months, spontaneous complaints, the commonest of which were nausea and vomiting, were made by 38% of patients receiving the combined and 39% receiving the separate formulations; 1% compared with 5%, respectively, complained that the tablets or capsules were too many, too large, difficult to swallow or that they stuck in the throat (p less than 0.05), and 32% compared with 45% regularly brought their own drink (usually milk or fruit juice) to the clinic to help them swallow their medicament (p less than 0.01). Only 14% of patients in each group missed one or more doses through default. Reported adverse reactions were mainly trivial, 4% of patients receiving the combined and 7% receiving the separate formulations having the administration of one or more drugs terminated. Thus, the differences between the combined and separate formulations showed a small advantage to the combined formulation in terms of acceptability to patients.


Machine Learning and Intelligent Diagnostics in Dental and Orofacial Pain Management: A Systematic Review.
The study explored the clinical influence, effectiveness, limitations, and human comparison outcomes of machine learning in diagnosing (1) dental diseases, (2) periodontal diseases, (3) trauma and neuralgias, (4) cysts and tumors, (5) glandular disorders, and (6) bone and temporomandibular joint as possible causes of dental and orofacial pain. Scopus, PubMed, and Web of Science (all databases) were searched by 2 reviewers until 29th October 2020. Articles were screened and narratively synthesized according to PRISMA-DTA guidelines based on predefined eligibility criteria. Articles that made direct reference test comparisons to human clinicians were evaluated using the MI-CLAIM checklist. The risk of bias was assessed by JBI-DTA critical appraisal, and certainty of the evidence was evaluated using the GRADE approach. Information regarding the quantification method of dental pain and disease, the conditional characteristics of both training and test data cohort in the machine learning, diagnostic outcomes, and diagnostic test comparisons with clinicians, where applicable, were extracted. 34 eligible articles were found for data synthesis, of which 8 articles made direct reference comparisons to human clinicians. 7 papers scored over 13 (out of the evaluated 15 points) in the MI-CLAIM approach with all papers scoring 5+ (out of 7) in JBI-DTA appraisals. GRADE approach revealed serious risks of bias and inconsistencies with most studies containing more positive cases than their true prevalence in order to facilitate machine learning. Patient-perceived symptoms and clinical history were generally found to be less reliable than radiographs or histology for training accurate machine learning models. A low agreement level between clinicians training the models was suggested to have a negative impact on the prediction accuracy. Reference comparisons found nonspecialized clinicians with less than 3 years of experience to be disadvantaged against trained models. Machine learning in dental and orofacial healthcare has shown respectable results in diagnosing diseases with symptomatic pain and with improved future iterations and can be used as a diagnostic aid in the clinics. The current review did not internally analyze the machine learning models and their respective algorithms, nor consider the confounding variables and factors responsible for shaping the orofacial disorders responsible for eliciting pain.


The regulation of cell-cell adhesion during epithelial-mesenchymal transition, motility and tumor progression.
Adherens junctions (AJs) are essential for the maintenance of epithelial homeostasis and a key factor in the regulation of cell migration and tumor progression. AJs maintain cell-cell adhesion by linking transmembrane proteins to the actin cytoskeleton. Additionally, they participate in recruitment of signaling receptors and cytoplasmic proteins to the membrane. During cellular invasion or migration, AJs are dynamically regulated and their composition modified to initiate changes in signaling pathways and cytoskeleton organization involved in cellular motility. Loss of E-cadherin, a key component of AJs, is characteristic of epithelial-mesenchymal-transition (EMT) and is associated with tumor cell invasion. We will review recent findings describing novel mechanisms involved in E-cadherin transcription regulation, endocytosis of E-cadherin and signaling associated with loss of AJs as well as reorganization of the AJ during EMT.


A detailed examination of substance P in pathologically graded cases of Huntington's disease.
Substance P concentrations have been found to be reduced in the basal ganglia in Huntington's disease (HD). In order to further examine this finding in the present study we measured substance P-like immunoreactivity (SPLI) in cases of HD which had been graded as to the severity of pathological changes in the striatum. Marked significant reductions of SPLI were found in all striatal nuclei which were significantly correlated with the percentage of neuronal loss in the varying pathologic grades. Similar changes were found in the projection sites of striatal substance P neurons, the globus pallidus interna and the substantia nigra. These changes are consistent with a loss of striatal substance P containing projection neurons in HD. Significant reductions in SPLI were also found in the external pallidum, bed nucleus of the stria terminalis and the subthalamic nucleus. Small significant increases in SPLI (20-30%) were found in 3 frontal cortical regions (Brodmann areas 6, 8 and 9). The finding of neurochemical changes in the subthalamic nucleus is of particular interest since lesions in this nucleus are known to result in chorea and therefore might contribute to the chorea which is a cardinal symptom of HD.


Detection of Helicobacter pylori in virtual slides requires high resolution digitalisation.
To diagnose Helicobacter pylori (HP) infection and its related mucosal injuries requires the histopathological analysis of gastric biopsies. The move from glass slides interpretation towards digital pathology implies technical choices to maintain the performances of histopathological diagnosis. The intra-rater agreement in assessing gastritis diagnostic criteria between glass slides, low resolution and high resolution digital slides in the subject of the present study. One hundred gastric biopsies were re-assessed by a single digestive pathologist on glass slides and digitalised slides at low resolution (ie, x20 magnification and single focus without z-stack) and high resolution (ie, x40 magnification with seven focus levels and z-stack) about the criteria of the updated Sydney system and the detection of HP. Inter-analyses agreement were very good (Kappa values>0.81) for every criteria but slightly inferior (ie, Kappa values<0.9) comparing glass slides interpretations with low resolution digital slides-based ones. Indeed, some HP infections were misdiagnosed using x20 magnification histochemical stained digitalised slides (p<0.05). At the opposite, anti-HP immunohistochemistry slides and/or x40 magnification digitalisation permitted to maintain almost perfect concordance in diagnosis (Kappa value>0.9). As mentioned in current guidelines, a high resolution x40 magnification digitalisation must be favoured in order to avoid some misdetection of microorganisms as HP.


Impact of IL28B and PNPLA3 polymorphisms on treatment outcomes in patients infected with genotype 6 hepatitis C virus.
Interleukin-28B (IL28B) and patatin-like phospholipase domain containing 3 (PNPLA3) gene polymorphisms are associated with hepatitis C virus (HCV) clearance and fatty liver, respectively. We aimed to test if their polymorphisms are associated with virologic responses in Chinese chronic hepatitis C (CHC) patients. This was a retrospective-prospective cohort study. Consecutive patients infected by genotype 1 and 6 HCV received antiviral therapy were included. Host IL-28B rs12979860/rs8099917 and PNPLA3 rs738409 genotype were tested. The primary outcome was sustained virologic response (sustained virologic response [SVR]: undetectable HCV RNA 24 weeks post-treatment). From 305 patients had positive antibody to HCV, 52 and 31 patients infected by genotype 1 and 6 HCV, respectively were recruited. Mean age was 58 ± 11 years; 70% were male. Mean baseline HCV RNA was 6.8 ± 2.7 log IU/ml. The SVR for patients infected by genotype 1 and 6 HCV was 67.3% and 90.3%, respectively. The proportions of IL28B genotypes were 78%, 21%, and 1% for TT/TG/GG at rs8099917, and 81%, 18%, and 1% for CC/TC/TT at rs12979860, respectively. The proportions of PNPLA3 rs738409 genotypes were 16%, 36%, and 48% for GG/GC/CC. IL28B genotype was significantly associated with SVR in patients infected by genotype 1 but not genotype 6 HCV, with 80% versus 38% of patients infected by genotype 1 achieved SVR carried TT versus TG/GG at rs8099917, respectively (P=0.003). PNPLA3 genotype was not associated with SVR. IL28B gene with rs8099917 T allele as an independent predictor of SVR in Chinese CHC patients infected by genotype 1 but not genotype 6 HCV.


Stress, appraisal and coping in HIV-infected African-American men and women.
With the development and utilization of the new multidrug therapies, many persons with HIV have experienced dramatic improvements in their physical condition, health self-appraisal, and psychological well-being. This treatment process has demonstrated the interrelationship of physical stress, perceived health status, and successful coping with HIV illness. The objective of this analysis is to assess the importance of perceived health for the level of mental distress in African-American men and women who have HIV disease. Interviews were completed with 255 HIV-infected black adults (age > or = 18) who were clients of AIDS care and referral centers in Tennessee. The results suggest that persons with a positive health appraisal are more effectively coping with the stresses of HIV illness. These observations have clinical implications for the care of African-Americans and others with HIV/AIDS.


Measurement of quality to improve care in sleep medicine.
The Board of Directors of the American Academy of Sleep Medicine (AASM) commissioned a Task Force to develop quality measures as part of its strategic plan to promote high quality patient-centered care. Among many potential dimensions of quality, the AASM requested Workgroups to develop outcome and process measures to aid in evaluating the quality of care of five common sleep disorders: restless legs syndrome, insomnia, narcolepsy, obstructive sleep apnea in adults, and obstructive sleep apnea in children. This paper describes the rationale, background, general methods development, and considerations in implementation for these sleep disorder quality measures. The Workgroup papers are published in this issue under the following titles: Quality Measures for the Care of Adult Patients with Restless Legs Syndrome, Quality Measures for the Care of Patients with Insomnia, Quality Measures for the Care of Patients with Narcolepsy, Quality Measures for the Care of Adult Patients with Obstructive Sleep Apnea, and Quality Measures for the Care of Pediatric Patients with Obstructive Sleep Apnea.


Cultural care of Thai immigrants in Uppsala: a study of transcultural nursing in Sweden.
The purpose of this study was to discover and describe the meanings and expressions of cultural care of a group of Thai immigrants in Sweden. Participants included 15 key informants and 24 general informants living in and around the town of Uppsala. The conceptual framework was provided by Leininger's theory of cultural care diversity and universality. Use was made of the ethnonursing method and the Sunrise Model in the search for multiple and related dimensions that influenced the generic and professional care practices of the Thai immigrants. Four major themes were formulated. Thus, care (a) means family and kinship relationships as expressed in daily life, (b) is expressed in traditional gender roles, (c) means religious beliefs as expressed in the Buddhist worship, and (d) means support of traditional health care practices. These themes support the cultural care theory and also confirm the Sunrise Model.


Preclinical models of congestive heart failure, advantages, and limitations for application in clinical practice.
Congestive heart failure (CHF) has increased over the years, in part because of recent progress in the management of chronic diseases, thus contributing to the maintenance of an increasingly aging population. CHF represents an unresolved health problem and therefore the establishment of animal models that recapitulates the complexity of CHF will become a critical element to be addressed, representing a serious challenge given the complexity of the pathogenesis of CHF itself, which is further compounded by methodological biases that depend on the animal species in use. Animal models of CHF have been developed in many different species, with different surgical procedures, all with promising results but, for the moment, unable to fully recapitulate the human disease. Large animal models often provide a more promising reality, with all the difficulties that their use entails, and which limit their performance to fewer laboratories, the costly of animal housing, animal handling, specialized facilities, skilled methodological training, and reproducibility as another important limiting factor when considering a valid animal model versus potentially better performing alternatives. In this review we will discuss the different animal models of CHF, their advantages and, above all, the limitations of each procedure with respect to effectiveness of results in terms of clinical application.


Brassinosteroids regulate grain filling in rice.
Genes controlling hormone levels have been used to increase grain yields in wheat (Triticum aestivum) and rice (Oryza sativa). We created transgenic rice plants expressing maize (Zea mays), rice, or Arabidopsis thaliana genes encoding sterol C-22 hydroxylases that control brassinosteroid (BR) hormone levels using a promoter that is active in only the stems, leaves, and roots. The transgenic plants produced more tillers and more seed than wild-type plants. The seed were heavier as well, especially the seed at the bases of the spikes that fill the least. These phenotypic changes brought about 15 to 44% increases in grain yield per plant relative to wild-type plants in greenhouse and field trials. Expression of the Arabidopsis C-22 hydroxylase in the embryos or endosperms themselves had no apparent effect on seed weight. These results suggested that BRs stimulate the flow of assimilate from the source to the sink. Microarray and photosynthesis analysis of transgenic plants revealed evidence of enhanced CO(2) assimilation, enlarged glucose pools in the flag leaves, and increased assimilation of glucose to starch in the seed. These results further suggested that BRs stimulate the flow of assimilate. Plants have not been bred directly for seed filling traits, suggesting that genes that control seed filling could be used to further increase grain yield in crop plants.


High-speed plasma measurements with a plasma impedance probe.
Plasma impedance probes (PIPs) are a type of RF probe that primarily measures electron density. This work introduces two advancements: a streamlined analytical model for interpreting PIP-monopole measurements and techniques for achieving ≥1 MHz time-resolved PIP measurements. The model's improvements include introducing sheath thickness as a measurement and providing a more accurate method for measuring electron density and damping. The model is validated by a quasi-static numerical simulation, which compares the simulation with measurements, identifies sources of error, and provides probe design criteria for minimizing uncertainty. The improved time resolution is achieved by introducing higher-frequency hardware, updated analysis algorithms, and a more rigorous approach to RF calibration. Finally, the new model and high-speed techniques are applied to two datasets: a 4 kHz plasma density oscillation resolved at 100 kHz with densities ranging between 2 × 1014 and 3 × 1015 m-3, and a 150 kHz oscillation resolved at 4 MHz with densities ranging between 4 × 1014 and 6 × 1014 m-3.


Syndromes presenting adducted thumb with/without clubfoot and Dundar syndrome.
Congenital adducted thumb has been called variously as congenital clasped thumb, thumb in palm deformity or flexion adduction deformity of the thumb. This condition can be an isolated anomaly or associated with several genetic disorders. The syndromes that include adducted thumb as a cardinal feature such as Dundar Syndrome are few in the literature. This syndrome is an autosomal-recessive very rare disorder characterized by typical facial appearance with dysmorphic features that includes wasted build, hyperextensible, thin and translucent skin with atrophic scarring, severe congenital contractures of fingers and thumbs, club feet, severe kyphoscoliosis, joint instability, muscular hypotonia, and ocular involvement. Heart, kidney, and/or intestinal defects can also be observed. Up to date the syndrome is described in few families in the literature. Here we discuss the syndromes that include adducted thumb as a cardinal feature and also the differential diagnosis of the Dundar Syndrome according to the literature.


[A perspective on living conditions: child and adolescent mortality in Manguinhos, Rio de Janeiro, Brazil].
Using data from the Mortality Information System, this paper investigates the deaths of children and adolescents in the Manguinhos neighborhood from 1996 to 2000, to determine the main characteristics and associate key mortality aspects with local living conditions. An outlying working-class or "suburban" neighborhood of the city of Rio de Janeiro where the main campus of the Oswaldo Cruz Foundation is located, Manguinhos consists of 12 "communities" characterized by poverty, social exclusion, drug traffic, and structural violence. In light of these factors and the cultural, social, and economic potential of the Manguinhos population, the authors begin with the theoretical/ practical premise that living conditions played a major role in the deaths of these young citizens. The study of these fatal events may therefore lead to the elucidation of issues and problems that must be included on the agendas and in forums involving both health promotion projects and the Municipal, State, and Federal governments so that they can be appropriately addressed in the scope of public policies.


Surgical treatment of scoliosis in osteogenesis imperfecta with cement-augmented pedicle screw instrumentation.
A retrospective study. To report the early postoperative results of scoliosis surgery in osteogenesis imperfecta (OI) patients utilizing all pedicle screw constructs and present a novel cementing technique to increase pedicle screw purchase in the osteoporotic OI spine. Scoliosis surgery utilizing hooks and wire systems have high complication rates in OI. Pedicle screw fixation systems have the biomechanical advantage of 3-column fixation, and cement augmentation of pedicle screws provides additional pull-out strength in the osteoporotic OI spine. The clinical and radiologic results of 10 consecutive OI patients treated with all pedicle screw instrumentation and fusion were retrospectively reviewed. The radiologic data included preoperative and postoperative major curve measurements: major curve Cobb angle, global coronal balance (GCB), apical vertebral translation (AVT), and the lowest instrumented vertebral (LIV) tilt. Operative findings included blood loss, surgery time, and additional procedures. All patients received intravenous pamidronate therapy preoperatively to increase bone mineral density. Ten patients with OI were operated on between 2005 and 2009. Seven had cement-augmented pedicle screw insertion at the proximal and distal foundations. The mean hospital stay was 10±7.5 days (range, 4-27 d) and the average follow-up period was 25.7±13.1 months (range, 14-50 mo). Mean preoperative and postoperative major Cobb angles were 83.7±23.8 and 40.3±14.6 degrees, respectively (48% correction; P<0.05). Mean preoperative and postoperative GCB deviations were 26.7±18.6 and 14.1±13.3 mm, respectively (P=0.097). Mean preoperative and postoperative AVTs were 69.3±29.1 and 29±12.2 mm, respectively (P<0.05). Preoperative and postoperative LIV tilts were 18.5±8.9 and 5.2±3.9 degrees, respectively (P<0.05). At the latest follow-up, the mean major curve Cobb angle was 37.7±13.1 degrees, the GCB deviation was 13.8±5.1 mm, the AVT was 31.7±13.3 mm, and the LIV tilt was 11.3±8.8 degrees. There was no difference between the early postoperative and the latest follow-up major curve Cobb angle, GCB deviation, AVT, or LIV tilt, indicating maintenance of correction. The mean blood loss was 23,75 mL (range, 800-45,00 mL). The mean operative time was 375.4 minutes (range, 262-491 min). The mean postoperative Scoliosis Research Society-22 patient-based outcome scores were 4.6±0.7 (out of 5). There were no instrumentation failures or permanent neurological deficits in this series. Pedicle screw instrumentation in OI scoliosis is safe and effective. Cement augmentation in these patients may help to increase the pedicle pull-out strength and decrease the screw failure rates, especially at the proximal and the distal ends of instrumentation.


Isolation and characterization of acyclovir-resistant mutants of herpes simplex virus.
Mutants of HSV which are resistant to acyclovir (acycloguanosine) have been isolated following serial passages of several herpes simplex virus (HSV) strains in the presence of the drug. The majority of the mutants isolated are defective in induction of thymidine kinase (TK) and this is consistent with the observation that independently isolated TK- viruses are naturally resistant to ACV. One mutant is described (SC16 R9C2) which is resistant in biochemically transformed cells which express HSV TK. This suggests that its resistance resides at a level other than TK. It is also resistant to phosphonoacetic acid, suggesting that the DNA polymerase locus may be involved. A further mutant is described [Cl (101) P2C5] which induces normal levels of TK, although the nature of resistance of this virus is not yet elucidated.