Cellular immune activation in children with acute dengue virus infections is modulated by apoptosis.
Apoptosis is an important modulator of cellular immune responses during systemic viral infections. Peripheral-blood mononuclear cell (PBMC) apoptosis and plasma soluble levels of CD95, a mediator of apoptosis, were determined in sequential samples from children participating in a prospective study of dengue virus (DV) infections. During the period of defervescence, levels of PBMC apoptosis were higher in children developing dengue hemorrhagic fever (DHF), the most severe form of illness, than in those with dengue fever (DF) and other, nondengue, febrile illnesses. CD8(+) T lymphocytes made up approximately half of the peak circulating apoptotic PBMCs in DHF and DF. Maximum plasma levels of soluble CD95 were also higher in children with DHF than in those with DF. The level of PBMC apoptosis correlated with dengue disease severity. Apoptosis appears to be involved in modulation of the innate and adaptive immune responses to DV infection and is likely involved in the evolution of immune responses in other viral hemorrhagic fevers.


Quantification of physical activity performed during US Army Basic Combat Training.
During US Army Basic Combat Training (BCT), graduation requirements, including physical readiness training (PRT), are standardized across training sites. However, there are concerns that the standardization may not be closely followed. Therefore, the purpose of this study was to measure and compare physical activity (PA) performed by recruits at 2 Army BCT sites. Twenty-four recruits per company from 11 companies (n=144 at Fort Jackson, SC; n=120 at Fort Sill, OK) wore an accelerometer and completed a daily PA log. The PA of one recruit from each company was recorded using an Army-developed direct observation tool (PAtracker). Amounts of time spent in various activity types, intensities, body positions, and in carrying external loads were obtained from the accelerometer, PA log, and PAtracker. Independent samples t tests were used to compare PA percentage time (%T) across training sites. Repeated measures analysis of variance was used to examine weekly differences in time spent in moderate to vigorous intensity PA during morning PRT. Physical activity was measured for 47 days at Fort Jackson and 44 days at Fort Sill. Differences in the percentage of time spent in various physical activities between the 2 sites ranged from 0.4% to 15.3% (2.0-93.7 minutes). At Fort Jackson, time spent in moderate to vigorous PA during PRT significantly increased each week for the first 4 to 6 weeks of BCT. No difference was observed in PAtracker data between the 2 training sites in the percentage of time recruits spent in calisthenics (3.9%±3.6% vs 3.8%±3.0%, P=.700), and only a small difference was observed in percentage of time recruits spent running (1.2%±1.7% vs 1.6%±2.0%, P=.037). Army recruits at the 2 BCT sites spent similar amounts of time in each PA variable, regardless of the training site and measurement method.


Our expedition in eight-membered ring compounds: from planar dehydrocyclooctenes to tub-shaped chiral tetraphenylenes.
Our research focus for almost forty years concerning eight-membered ring compounds is herein summarized. The design and preparation of these structurally unique compounds featuring a central cyclooctene ring are described in the context of their planar or tub-shaped conformations. Furthermore, their intrinsic properties and potential applications are also presented.


[Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefecture (2010-2011)].
We investigated genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, the serotypes and the susceptibility to antibacterial agents against 258 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between January 2010 and March 2011. These results were compared with those against 377 strains of S. pneumoniae isolated in 2008-2009. The number of genotype penicillin-susceptible S. pneumoniae (gPSSP) with 3 normal PBP genes, genotype penicillin-intermediate S. pneumoniae (gPISP) with 1 or 2 normal PBP genes and genotype penicillin-resistant S. pneumoniae (gPRSP) with 3 abnormal genes was 11 (4.3%), 135 (52.3%) and 112 (43.4%) strains, respectively. The isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 17 (6.6%), 65 (25.2%), 143 (55.4%) and 33 (12.8%). The prevalent pneumococcal serotypes isolated from children were type 19F (18.2%), following by type 6A and 15 (11.7%). The potential coverage of pneumococcal conjugate vaccine (PCV7) was 43.8%. The prevalent pneumococcal serotypes isolated from adults were high in order of type 19F (12.8%), type 6A, 3 and 11 (10.3%), excepting non-typable strains (17.9%), and from elderly persons were type 6B (23.2%) and type 3 (13.4%). The MIC90 of each antibacterial agents was as follows; 0.0625 microg/mL for garenoxacin, 0.125 microg/mL for panipenem, 0.25 microg/mL for imipenem, doripenem, tosufloxacin, 0.5 microg/mL for cefditoren, meropenem, moxifloxacin, 1 microg/mL for amoxicillin, clavulanic acid/amoxicillin, cefteram, cefcapene, ceftriaxone, 2 microg/mL for benzylpenicillin, piperacillin, tazobactam/ piperacillin, pazufloxacin, levofloxacin, 4 microg/mL for cefdinir, flomoxef, 16 microg/mL for minocycline, > 64 microg/mL for clarithromycin, azithromycin and these MIC90s were about the same as those in 2008-2009.


[Effectiveness of 23-valent penumococcal polysaccharide and split-virus influenza vaccines to prevent respiratory diseases for elderly people].
To assess the effectiveness of 23-valent penumococcal polysaccharide vaccine (PenV23) and Split-Virus influenza vaccine (InfV-B) for preventing upper respiratory diseases in the aging population. 151 people aged > or =60 years who inoculated the PenV23 and InfV-B vaccines at clinics during 2005 as trial group, and 188 people aged > or =65 years didn't administered as control guoup from some comminity. On base line survey, both guoups were followed up two years after vaccination about incidence, hospitalization rate, treatment and direct medical cost. The vaccine of PenV23 and InfV-B vaccination in upper respiratory tract infection was 60% [(odds ratio OR) = 0.35, 95% CI (0.153-0.794)] and the difference between the 2 groups was significant (P = 0.009). The people had recevied both vaccines, the mean duration of hospitalization in vaccinated vs unvaccinated control peple was 20.00 days: 24.19 days (t = 5.82, P < 0.001). Benefit-cost ratio was 4.03, and the net benefit was Yen 177994.86. Incidence of influenza-like illness for the elderly people in the community decreased by administered PenV23 ahd InfV-B vaccine. This vaccines had high cost-benefits.


Comparison of transcatheter and surgical treatment of paravalvular leak: Results from a 5-year follow-up study.
This study aimed to compare the efficacy and safety of two different treatments of paravalvular leak (PVL). PVL is a common complication after surgical valve replacement. Re-operation is associated with high mortality, morbidity, and risk of re-leak. Catheter-based repair has emerged as a promising new therapy. Eighty-seven consecutive patients with symptomatic PVL received either transcatheter (n = 46) or surgical (n = 41) treatment at Shanghai Chest Hospital between January 2009 and December 2015.The procedural and clinical success rates were similar between the transcatheter group and the surgical group (82.6 vs. 90.2%; P = 0.30; and 69.5 vs. 73.0%; P = 0.71, respectively). There were fewer in-hospital total major adverse events in the transcatheter group (56.09 vs. 17.39%; P < 0.001), and transcatheter repair was more cost-effective, with fewer blood transfusions, shorter procedure durations, shorter hospital stays, and less expenditure. However, there were six cases of hemolysis aggravation in the transcatheter group (13.04%). The 5-year overall survival rates after transcatheter and surgical repair were 74.39 and 71.95% (P = 0.45), respectively, and the cardiac-related survival rates were 84.08 and 74.72% (P = 0.19), respectively. Transcatheter and surgical repairs are both effective treatments for selected patients with PVL. And, transcatheter closure seems to be safer and more cost-effective. Nonetheless, this new treatment may be risky for post-procedure hemolysis when unsuitable devices are used.


Nucleotide sequence of the Serratia marcescens threonine operon and analysis of the threonine operon mutations which alter feedback inhibition of both aspartokinase I and homoserine dehydrogenase I.
The nucleotide sequence of the Serratia marcescens threonine operon (thrA1A2BC) was determined. Three long open reading frames were identified; these open reading frames code for aspartokinase I (AKI)-homoserine dehydrogenase I (HDI), homoserine kinase, and threonine synthase, in that order. The predicted amino acid sequences of these enzymes were similar to the amino acid sequences of the corresponding enzymes in Escherichia coli. The AKI-HDI protein is apparently a tetramer composed of monomer polypeptides that are 819 amino acids long. A deletion analysis revealed that the central and C-terminal region was responsible for threonine-resistant HDI activity, a monomeric fragment extending from the N terminus to residue 306 was responsible for threonine-resistant AKI activity, and an N-terminal portion containing 468 residues was responsible for threonine-sensitive AKI activity. The thrA(1)1A(2)1 and thrA(1)5A(2)5 mutations of threonine-excreting strains HNr21 and TLr156, which result in the loss of threonine-mediated feedback inhibition of both AKI activity and HDI activity, cause single amino acid substitutions (Gly to Asp at position 330 and Ser to Phe at position 352, respectively) in the central region of the AKI-HDI protein. The thrA1+A(2)2 mutation of strain HNr59, which results in a threonine-sensitive AKI and a threonine-resistant HDI, also causes a single amino acid substitution (Ala to Thr at position 479).


Plasma amino acid analyses in two cases of maple syrup urine disease.
Maple syrup urine disease is a rare inborn error of metabolism, characterized by elevated plasma levels of branched chain amino acids and urinary excretion of branched chain keto acids. Plasma amino acid levels in two subjects were followed by deproteinizing plasma, derivatizing the free amino acids with phenylisothiocyanate, and analysis by HPLC. The results indicate that valine, leucine and isoleucine are elevated in Maple syrup urine disease, and that leucine remains high even after dietary treatment.


The Monocyte-Derived Exosomal CLMAT3 Activates the CtBP2-p300-NF-κB Transcriptional Complex to Induce Proinflammatory Cytokines in ALI.
Monocytes and macrophages are the two major cell types involved in innate immunity. Exosomes act as signaling molecules to regulate cell-to-cell communication by releasing proteins, mRNAs, microRNAs (miRNAs), and long noncoding RNAs (lncRNAs). However, it is still unclear whether monocyte-derived exosomes are involved in the communication between monocytes and macrophages. In this study, we analyzed the differentially expressed lncRNA profiles in monocytes isolated from blood samples of healthy controls and acute lung injury (ALI) patients. We focused our study on investigating the signaling downstream of CLMAT3 (colorectal liver metastasis-associated transcript 3), a lncRNA that regulated proinflammatory cytokine genes. We revealed that CLMAT3 specifically targeted CtBP2 (C-terminal binding protein 2) and repressed its expression. Elevated CtBP2 acted as a coactivator to assemble a transcriptional complex with histone acetyltransferase p300 and NF-κB (nuclear factor κB) subunits. In vitro coculture and in vivo injection of ALI monocyte-derived exosomes increased the production of proinflammatory cytokines. Importantly, the administration of two CtBP2 inhibitors, NSC95397 and MTOB, could significantly reverse CtBP2-mediated transactivation. Collectively, our results support a model in which monocyte-derived exosomal CLMAT3 activates the CtBP2-p300-NF-κB complex to induce proinflammatory cytokines, thus contributing to the pathogenesis of ALI.


Investigation of an outbreak of monkeypox in an area occupied by armed groups, Central African Republic.
Monkeypox is a zoonosis caused by an Orthopoxvirus of the Poxviridae family. Human infections are often severe and are a public health problem. We conducted a monkeypox outbreak investigation of suspected case patients in five villages of the Alindao-Mingala Health District following blood sample confirmation of the virus by the Institut Pasteur in Bangui. We aimed to determine disease characteristics, to describe the context and the risk factors, and to measure the incidence and case fatality. Patients were reported in the villages of Rehou 4, 5, Dalakere 1, Kongbo, and Pavika from August to October 2016. Data was collected on individual records when interviewing patients or parents. A total of 26 patients were identified. The˂10 years and 21-30 years age groups were the most affected. The overall attack rate was 5 per 1000 inhabitants and the case fatality was 7.7%. Young age and the absence of smallpox vaccination were associated with severe presentations in 87.5% of cases. The annual number of monkeypox outbreaks increases in the Central African Republic with severe presentations and a high case fatality especially in children. Reinforcing the surveillance and characterization of circulating strains will provide information on the need for vaccine production.


Holography and string dynamics in time-dependent backgrounds.
We study the dynamics of D-branes in a smooth time-dependent background. The theory on the branes is a time-dependent noncommutative field theory. We find the metric and fluxes that determine the dual holographic closed string theory. This provides a concrete example of holography in a cosmological setting.


Effects of ambient relative humidity and solvent properties on the electrospinning of pure hyaluronic acid nanofibers.
Nanofibers exist ubiquitously in natural extracellular matrix (ECM) of all kinds of human tissues forming hydrated interwoven network. Electrospinning nanotechnology has been proven to be a powerful technique to fabricate controllable nanofibers mimicking the natural ECM structures. Hyaluronic acid (HA), as a critical component of natural ECM, has been widely used in tissue engineering and regenerative medicine. In this study, pure HA nanofibers with average diameter of 33 +/- 5 nm, 59 +/- 12 nm, 79 +/- 12 nm and 113 +/- 19 nm were successfully prepared using different electrospinning parameters. The effect of the ambient relative humidity on HA electrospinnability was investigated for the first time in detail, which was proven to be one of the most important factors to control the morphology of HA nanofibers beside the solution properties. A critical value of humidity for a defined HA solution was observed, only below which HA nanofibers with similar diameters and morphologies could be successfully obtained. When the ambient relative humidity was higher than the critical value, the HA nanofibers started dissolving at the cross points and even fused together forming a spreading layer. Moreover, only a small amount of N, N-Dimethylformamide (DMF) was found to be required to promote the electrospinnability of HA solution by mixing with water as solvents. With the increase in the DMF content, the surface tension of the solution decreased significantly, which was thought to be benefit for the stable Taylor cone and fluid jet formation in electrospinning. At the same time, it should be noted that the conductivity of the solution also decreased with the increase of DMF content in the solution, which was believed to be responsible for the increasing diameters of HA nanofibers corresponding to higher DMF content. Controllable HA nanofibers with diameter below 100 nm have great promising for developing novel nanobiomaterials applied in tissue engineering and regenerative medicine.


COVID-19 survivors: Multi-disciplinary efforts in psychiatry and medical humanities for long-term realignment.
The coronavirus disease 2019 pandemic represents an enduring transformation in health care and education with the advancement of smart universities, telehealth, adaptive research protocols, personalized medicine, and self-controlled or artificial intelligence-controlled learning. These changes, of course, also cover mental health and long-term realignment of coronavirus disease 2019 survivors. Fatigue or anxiety, as the most prominent psychiatric "long coronavirus disease 2019" symptoms, need a theory-based and empirically-sound procedure that would help us grasp the complexity of the condition in research and treatment. Considering the systemic character of the condition, such strategies have to take the whole individual and their sociocultural context into consideration. Still, at the moment, attempts to build an integrative framework for providing meaning and understanding for the patients of how to cope with anxiety when they are confronted with empirically reduced parameters (e.g., severe acute respiratory syndrome coronavirus type 2) or biomarkers (e.g., the FK506 binding protein 5) are rare. In this context, multidisciplinary efforts are necessary. We therefore join in a plea for an establishment of 'translational medical humanities' that would allow a more straightforward intervention of humanities (e.g., the importance of the therapist variable, continuity, the social environment, etc) into the disciplinary, medial, political, and popular cultural debates around health, health-care provision, research (e.g., computer scientists for simulation studies), and wellbeing.


Di George anomaly associated with a de novo Y;22 translocation resulting in monosomy del(22)(q11.2).
We report on an infant, born to a diabetic mother, who presented with hypocalcemia and congenital heart disease, presurgically diagnosed by echocardiography as truncus arteriosus type I. Cytogenetic analysis showed a 45,X,-Y,-22,+der-(Y)t(Y;22) (p11.3q11.2) chromosome abnormality with del(22)(q11.2). Parental chromosomes were normal. Autopsy showed persistent truncus arteriosus type II and thymic aplasia consistent with DiGeorge anomaly. This report adds to the existing literature demonstrating an association between DiGeorge anomaly and monosomy 22q11.


DUETS (Dallas UtErus Transplant Study): The role of imaging in uterus transplantation.
Uterus transplantation is rapidly becoming a viable clinical option for women with uterine-factor infertility and a desire for parenthood. Radiological imaging plays a central role in selecting the optimal living donors for uterus transplantation and serves to exclude any pathology and evaluate the uterine vasculature. The latter is the most important variable in the ultimate technical success of the uterus transplant. In this first report of imaging in the setting of uterus transplantation, we report our experience with living-donor selection, and the evolution of the imaging techniques that ultimately allowed a significant improvement in donor selection and transplant outcome. We also suggest a framework for preoperative imaging in uterus transplantation. Between 2016 and 2018, 27 potential living donors were evaluated by imaging prior to uterine donation for uterus transplantation. Predonation imaging included a screening chest radiograph, dual-phase computed tomography (CT) angiograms of the abdomen and pelvis in the arterial and venous phases and pelvic sonography with Doppler. Seventeen potential donors additionally underwent multiphasic pelvic MR angiograms. The imaging performed was meant to display features of the vascular anatomy relevant for uterus transplantation. Out of the 27 potential live donors who were evaluated by imaging, 9 eventually donated their uterus for transplantation. The most frequent reason for exclusion was suboptimal quality of the vessels (33%), including small uterine arteries, the presence of atherosclerosis or small size/poor quality of the uterine or utero-ovarian veins, or both. The next most common reason was voluntary patient withdrawal or failure to complete the evaluation process (28%). Three potential donors (16.6%) were rejected for uterine factors, fibroids, and/or adenomyosis. Other reasons for donor rejection included ABO incompatibility and unfavorable psychological evaluation. Diagnostic imaging plays a crucial role in selecting appropriate potential donors, screening prospective recipients, planning the graft procedure, and following up on any graft or nongraft-related complications in both the donor and recipient after the transplantation procedure is performed. Contrast-enhanced CT and MR angiographies have complementary roles, especially when evaluating the donor for adequacy of the arterial and venous supply to the uterine graft and the experience gained from our series indicates that the inclusion of both modalities contributed directly to successful uterus transplant graft survival by selecting patients with favorable arterial and venous vasculature.


Long-term treatment with a phosphodiesterase type 5 inhibitor improves pulmonary hypertension secondary to heart failure through enhancing the natriuretic peptides-cGMP pathway.
In advanced heart failure (HF), the compensatory pulmonary vasodilation is attenuated due to the relative insufficiency of cGMP despite increased secretion of natriuretic peptides (NPs). Phosphodiesterase type 5 (PDE5) inhibitors prevent cGMP degradation, and thus may potentiate the effect of the NPs-cGMP pathway. We orally administered a specific PDE5 inhibitor, T-1032 (1 mg/kg; twice a day, n = 7) or placebo (n = 7) for 2 weeks in dogs with HF induced by rapid pacing (270 bpm, 3 weeks) and examined the plasma levels of atrial natriuretic peptide (ANP), cGMP, and hemodynamic parameters. We also examined the hemodynamic changes after injection of a specific NPs receptor antagonist, HS-142-1 (3 mg/kg), under treatment with T-1032. T-1032 significantly increased plasma cGMP levels compared with the vehicle group despite low plasma ANP levels associated with improvement in cardiopulmonary hemodynamics. HS-142-1 significantly decreased plasma cGMP levels in both groups, whereas it did not change all hemodynamic parameters in the vehicle group. In contrast, in the T-1032 group, HS-142-1 significantly increased pulmonary arterial pressure and pulmonary vascular resistance. These results indicated that long-term treatment with a PDE5 inhibitor improved pulmonary hypertension secondary to HF and the NPs-cGMP pathway contributed to this therapeutic effect.


Is grand multiparity a risk factor for osteoporosis in postmenopausal women of lower socioeconomic status?
We aimed to determine whether grand multiparity is a risk factor for osteoporosis among postmenopausal women of lower socioeconomic status. We conducted a single center study between February 2012 and February 2013 on 50-60 year old postmenopausal women of lower socioeconomic status without a history of medical disease. Women with a body mass index (BMI) between 20 and 25 were included in the study. The grand multiparous group (group A) consisted of 38 women with 10 or more deliveries. Women with a history of three or fewer deliveries composed the control group (group B). Dual-energy x-ray absorptiometry was used to measure the bone mineral density (BMD) of the proximal femur neck and lumbar spine (L1-L4). The mean ages of groups A and B were found to be 54.3 ± 2.5 and 53.1 ± 2.7 years, respectively. Average parity in groups A and B was 11.1 ± 1.7 and 2.4 ± 0.7, respectively. Time since the onset of menopause was 3.6 ± 2.7 years in group A and 6.0 ± 2.9 in group B. The prevalence of osteoporosis was similar in both groups (71.1%-81.4%, p = 0.273). We found that grand multiparity was an ineffective indicator of either femoral or lumbar osteoporosis (p = 0.87 and p = 0.26), but osteoporosis five years after the onset of menopause was found to be significantly higher (p = 0.02). The duration of menopause is an independent risk factor of osteoporosis. However, the number of pregnancies is neither a determinant nor a protective factor for osteoporosis in postmenopausal women coming from a low socioeconomic background.


Epithelial cell invasiveness of non-enteropathogenic serotypes of Escherichia coli.
Current evidence suggests that enteropathogenic Escherichia coli (EPEC) of traditional serotypes possess a three-stage pathogenesis: localised adherence (LA), to, attachment-effacement (AE) of, and penetration of, enterocytes, all of which can be reproduced in tissue culture models in vitro. Three E. coli isolates of non-traditional serotypes (02:H2, 02:H25 and 015:H2) isolated from children with diarrhoea were previously shown to be positive for LA and AE activities in laboratory models. In the present study, they were, in addition, shown to be positive for invasion of a HEp-2 cell monolayer. These findings further establish the pathogenicity of non-traditional serotypes of E. coli and their role in the causation of diarrhoea.


Trophoblast Differentiation Affects Crucial Nutritive Functions of Placental Membrane Transporters.
Cytotrophoblasts are progenitor cells that proliferate and fuse to form the multinucleated syncytiotrophoblast layer, implicated in placental endocrine and transport functions. While membrane transporters play a critical role in the distribution of nutrients, hormones, and xenobiotics at the maternal-fetal interface, their selectivity to the syncytiotrophoblast layer is poorly characterized. We aimed to evaluate the regulation of placental transporters in response to trophoblast differentiation in vitro. Experiments were carried out in isolated primary human trophoblast cells before and after syncytialization. Gene expression of six molecular markers and thirty membrane transporters was investigated by qPCR analysis. Subsequently, functional expression was evaluated for proteins involved in the transplacental transfer of essential nutrients i.e., cholesterol (ABCA1, ABCG1), glucose (SLC2A1), leucine (SLC3A2, SLC7A5), and iron (transferrin receptor, TfR1). We identified that human chorionic gonadotropin, placental lactogen, endoglin, and cadherin-11 serve as optimal gene markers for the syncytialization process. We showed that trophoblast differentiation was associated with differential gene expression (mostly up-regulation) of several nutrient and drug transporters. Further, we revealed enhanced protein expression and activity of ABCG1, SLC3A2, SLC7A5, and TfR1 in syncytialized cells, with ABCA1 and GLUT1 displaying no change. Taken together, these results indicate that the syncytiotrophoblast has a dominant role in transporting essential nutrients cholesterol, leucine, and iron. Nonetheless, we present evidence that the cytotrophoblast cells may also be linked to transport functions that could be critical for the cell fusion processes. Our findings collectively yield new insights into the cellular functions associated with or altered by the trophoblast fusion. Importantly, defective syncytialization could lead to nutrient transfer imbalance, ultimately compromising fetal development and programming.


Transactivation specificity is conserved among p53 family proteins and depends on a response element sequence code.
Structural and biochemical studies have demonstrated that p73, p63 and p53 recognize DNA with identical amino acids and similar binding affinity. Here, measuring transactivation activity for a large number of response elements (REs) in yeast and human cell lines, we show that p53 family proteins also have overlapping transactivation profiles. We identified mutations at conserved amino acids of loops L1 and L3 in the DNA-binding domain that tune the transactivation potential nearly equally in p73, p63 and p53. For example, the mutant S139F in p73 has higher transactivation potential towards selected REs, enhanced DNA-binding cooperativity in vitro and a flexible loop L1 as seen in the crystal structure of the protein-DNA complex. By studying, how variations in the RE sequence affect transactivation specificity, we discovered a RE-transactivation code that predicts enhanced transactivation; this correlation is stronger for promoters of genes associated with apoptosis.


Composition of the salivary microflora during habitual consumption of fluoridated milk.
The aim was to evaluate the effect of habitual consumption of fluoridated milk on the composition of the salivary microflora. The study group comprised 20 healthy schoolchildren and young adults with a mean age of 13.6 years and the investigation had a randomized double-blind crossover design with a washout period of 1 month. After professional tooth-cleaning at baseline, the subjects were supplied with either fluoridated (250 mL, 5 ppm F) or non-fluoridated milk for one daily intake during a period of 4 weeks. Salivary samples were collected immediately before tooth-cleaning and after 1, 2, and 4 weeks, respectively. The samples were immediately cultivated for total viable counts, oral streptococci, mutans streptococci, lactobacilli, and actinomyces spp. Bacterial counts were logarithmically transformed before statistical evaluation using ANOVA. No significant alterations of the salivary microflora were found during any of the milk regimens compared with baseline. There was a slight reduction in the proportion of mutans streptococci after 2 and 4 weeks during consumption with fluoridated milk but the difference failed to reach statistical significance. In conclusion, this study was unable to disclose any significant alteration of the composition of the salivary microflora following daily intake of fluoridated milk.


Cytosolic and membrane-associated proteins involved in the recruitment of AP-1 adaptors onto the trans-Golgi network.
The AP-1 adaptor complex is recruited from the cytosol onto the trans-Golgi network membrane, where it co-assembles with clathrin into a coat that drives vesicle budding. The GTPase ARF1 has been shown to be required for AP-1 recruitment, and here we demonstrate that we can reconstitute full GTPgammaS-dependent recruitment of adaptors onto an enriched trans-Golgi network membrane fraction by adding purified AP-1 and recombinant myristylated ARF1, indicating that these are the only soluble proteins required for binding. To identify some of the membrane proteins involved in recruitment, we have incubated permeabilized metabolically labeled cells with cytosol under conditions that promote adaptor binding, then cross-linked the samples with 3,3'dithiobis(sulfosuccinimidylproprionate), denatured by boiling in SDS, and immunoprecipitated with antibodies against the various subunits. Under these conditions, the adaptor subunits co-precipitate not only with each other and with clathrin, but also with three novel proteins: p75, which is specifically cross-linked to gamma-adaptin; p80, which is specifically cross-linked to beta'-adaptin; and p60, which is specifically cross-linked to AP47. These proteins are all candidates for components of the adaptor docking site on the trans-Golgi network membrane.


Definition and risk factors of rapidly declining residual renal function in peritoneal dialysis: an observational study.
It is critical to preserve residual renal function (RRF) in peritoneal dialysis (PD), as RRF is associated with lower morbidity and mortality. There is no uniform definition of RRF, and rapidly declining RRF has rarely been studied and predominately limited to single factor analysis and not corrected for lead-time bias. An observational study in 71 incident PD patients. RRF was defined as urine output (UO) ≥500 ml/day and renal glomerular filtration rate (rGFR) ≥2 ml/min/1.73 m(2), rapid declining RRF as UO <500 ml/day and rGFR <2 ml/min/1.73 m(2) occurring within 6 months which were separately evaluated. Independent risk factors associated with rapid RRF decline were identified while correcting for lead-time bias. RRF declined rapidly by both definitions in 65% patients 2.5 years after PD start. Both definitions of RRF decline were consistent in 96%. Nephrotoxic drugs, renal transplant failure and absent angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB) were independent risk factors associated with rapidly declining RRF defined both by definitions, intravascular radiocontrast additionally for UO decline. Most PD patients demonstrated rapid RRF decline, independent of its definition. Both definitions are highly consistent and interchangeable. Nephrotoxic drugs and radiocontrast were identified as risk factors of acute, absent ACEI or ARB, and renal transplant failure of chronic renal injury.


Another discontinuous epitope on glycoprotein gp120 that is important in human immunodeficiency virus type 1 neutralization is identified by a monoclonal antibody.
To define the domains in the envelope glycoprotein important for antibody neutralization of the human immunodeficiency virus type 1 (HIV-1), monoclonal antibodies (mAbs) were generated by immunizing mice with purified glycoprotein gp120 of the IIIB isolate. One mAb, G3-4, reacted with the gp120 of homologous (IIIB) and heterologous (RF) isolates. In addition, mAb G3-4 efficiently neutralized both IIIB and RF viruses in vitro, as well as four of nine primary HIV-1 isolates. In competition immunoassays, mAb G3-4 and soluble CD4 were found to inhibit one another in binding to gp120. However, no competition was seen between mAb G3-4 and mAbs directed to the third variable region or the fourth conserved region of gp120. In particular, mAb G3-4 did not compete with our human mAb 15e, which identifies a discontinuous epitope on gp120 involved in group-specific neutralization of HIV-1 and in gp120-CD4 binding. Epitope-mapping studies on mAb G3-4 with synthetic or unglycosylated recombinant peptides were negative, suggesting that its epitope may be discontinuous. Indeed, this hypothesis was confirmed by showing the loss of mAb G3-4 serologic reactivity when gp120 was first denatured. We conclude that the site recognized by mAb G3-4 represents another discontinuous epitope on gp120 important for neutralization of HIV-1.


Assessment of left atrial mechanics in patients with atrial fibrillation: comparison between two-dimensional speckle-based strain and velocity vector imaging.
Two-dimensional (2D) speckle tracking-derived left atrial (LA) strain (ε) facilitates comprehensive evaluation of LA contractile, reservoir, and conduit function; however, its dependence on the individual software used for assessment has not been evaluated. The aim of this study was to compare LA ε derived from two different speckle-tracking software technologies, Velocity Vector Imaging (VVI) and 2D speckle-tracking echocardiography (STE). VVI-derived and 2D STE-derived global longitudinal LA ε and ε rate (SR) were directly compared in 127 patients (mean age, 62 ± 10 years) with atrial fibrillation. Peak negative, peak positive, and total ε (corresponding to LA contractile, conduit, and reservoir function) were measured during sinus rhythm. Late negative (LA contraction), peak positive (left ventricular systole), and early negative (left ventricular early diastole) SR were also measured. The measurement of LA ε and SR by both software was feasible in high proportions of patients (93% with VVI and 93% with 2D STE). The average analysis of ε(negative) was -7.24 ± 3.87% by VVI and -7.30 ± 3.37% by 2D STE (P = .84). The average analysis of ε(positive) was 14.52 ± 5.82% by VVI and 10.74 ± 4.51% by 2D STE (P < .01). The average analysis of ε(total) was 21.76 ± 7.39% by VVI and 18.04 ± 5.98% by 2D STE (P < .01). VVI-derived and 2D STE-derived ε(positive), ε(negative), and ε(total) had good correlations with one another (R = 0.79, R = 0.75, and R = 0.80), with low mean differences. Late negative, peak positive, and early negative SR were correlated less well (R = 0.78, R = 0.71, and R = 0.67). LA ε measurement using both VVI and 2D STE is feasible in a large proportion of patients in clinical practice. VVI and 2D STE provide comparable LA ε and SR measurements for LA contractile function.


Regulation of ovarian cholesterol metabolism: control of 3-hydroxy-3-methylglutaryl coenzyme A reductase and acyl coenzyme A:cholesterol acyltransferase.
Activities of enzymes involved in cholesterol metabolism were measured in ovaries of PMS-hCG-primed immature rats. Microsomal 3-hydroxy-3-methyglutaryl coenzyme A (HMG-CoA) reductase, a rate-limiting enzyme in sterol synthesis, was low during the period of maximal steroid production and cholesteryl ester storage. Acyl CoA:cholesterol acyltransferase (ACAT), the microsomal enzyme catalyzing sterol esterification and cytosolic cholesteryl ester hydrolase and mitochondrial cholesterol side chain cleavage activities, paralleled storage of sterol esters and levels of plasma progesterone, being highest on day 7 post hCG. Further experiments demonstrated that HMG-CoA reductase and ACAT are regulated by different mechanisms than the sterol esterase and cholesterol side chain cleavage system. When blood sterol levels were lowered by 4-aminopyrazolo (3,4-d)pyrimidine, plasma progestin concentrations fell and ovaries failed to accumulate sterol esters. HMG-CoA reductase increased more than 10-fold as a result of this treatment, while ACAT was 15% of that measured in controls. In contrast, cholesteryl esterase and the cholesterol side chain cleavage system were not affected. Intravenous infusion of human lipoproteins reversed the effects of 4-aminopyrazolo (3,4-d)pyrimidine. Raising blood levels up to 6-fold with either cholesterol-supplemented diets or intravenous injections of rat lipoproteins had no effect upon ovarian sterol metabolism, suggesting that the process by which blood cholesterol is utilized is saturated at normal blood sterol levels. Intravenous injection of LH on day 7 post hCG increased HMG-CoA reductase and decreased ACAT activity in addition to depleting cholesteryl esters. Prior treatment of rats with aminoglutethimide prevented the effects of LH on both HMG-CoA reductase and ACAT, indicating that changes in these enzymes were a consequence of the steroidogenic stimulus imposed by LH. Treatment with aminoglutethimide alone on day 7 post hCG did not further depress the already low HMG-CoA reductase activity, but ACAT and cholesterol ester storage were stimulated while sterol esterase activity was not altered. Aminoglutethimide also produced elevated ACAT activity and sterol ester storage in animals treated on day 2 post hCG, a time when there is normally little sterol ester accumulation. We conclude that HMG-CoA reductase and ACAT are regulated, in a reciprocal fashion, by ovarian cholesterol balance. When sufficient cholesterol is available, HMG-CoA reductase is suppressed and ACAT increases to facilitate esterification of sterol in excess of cell needs. When exogenous sterol supplies cannot meet ovarian demands, HMG-CoA reductase rises, and there is a concomitant decline in ACAT. Thus, gonadotropic control over these enzymes may be exerted, in part, through modulation of the supply of cholesterol to the ovary and/or through regulation of the rate of sterol utilization for hormone synthesis.


The role of activity in development of the visual system.
Neuronal activity is important for both the initial formation and the subsequent refinement of anatomical and physiological features of the mammalian visual system. Here we examine recent evidence concerning the role that spontaneous activity plays in axonal segregation, both of retinogeniculate afferents into eye-specific layers and of geniculocortical afferents into ocular dominance bands. We also assess the role of activity in the generation and plasticity of orientation selectivity in the primary visual cortex. Finally, we review recent challenges to textbook views on how inputs representing the two eyes interact during the critical period of visual cortical plasticity.


Mutational analysis of a conserved motif of Agrobacterium tumefaciens VirD2.
The VirD2 polypeptide from Agrobacterium tumefaciens, in the presence of VirD1, introduces a site- and strand-specific nick at the T-DNA borders. A similar reaction at the origin of transfer (oriT) of plasmids is essential for plasmid transfer by bacterial conjugation. A comparison of protein sequences of VirD2 and its functional homologs in bacterial conjugation and in rolling circle replication revealed that they share a conserved 14 residue segment, HxDxxx(P/u)HuHuuux [residues 126-139 of VirD2; Ilyina, T.V. and Koonin, E.V. (1992) Nucleic Acids Res. 20, 3279-3285]. A mutational approach was used to test the role of these residues in the endonuclease activity of VirD2. The results demonstrated that the two invariant histidine residues (H133 and H135) are essential for activity. Mutations at three sites, histidine 126, aspartic acid 128 and aspartic acid 130, that are conserved in a subfamily of the plasmid mobilization proteins, led to the loss of VirD2 activity. Aspartic acid at position 130, could be substituted with glutamic acid and to a much lesser extent, with tyrosine. In contrast, another conserved residue, asparagine 139, tolerated many different amino acid substitutions. The non-conserved residues, arginine 129, proline 132 and leucine 134, were also found to be important for function. Isolation of null mutations that map throughout this conserved domain confirm the hypothesis that this region is essential for function.


Effect of mercury on general and reproductive health of largemouth bass (Micropterus salmoides) from three lakes in New Jersey.
The influence of mercury on the general and reproductive health of wild fish populations has not been well studied. Therefore, a variety of health and reproductive indicators were measured in male largemouth bass (Micropterus salmoides) collected from three bodies of water in New Jersey: Assunpink Lake, Manasquan Reservoir, and Atlantic City Reservoir. The mean mercury content in fish muscle from Assunpink Lake was 0.30 microg/g; from Manasquan Reservoir, 1.23 microg/g; and from Atlantic City Reservoir, 5.42 microg/g. Body weight, length, condition factor, and gonadosomatic index were similar for all three lakes. Furthermore, there was no significant relationship between muscle mercury content and adrenocortical function, indicated by interrenal nuclear diameter and serum cortisol levels following stress. Bass from the Atlantic City Reservoir had a slightly lower, although significant, liver somatic index than bass from the two other lakes. A significant, positive correlation between 11-ketotestosterone serum concentrations and mercury muscle content was detected, although no significant relationship between testosterone serum concentrations and mercury muscle content was found. The findings of this study suggest that, while elevated levels of mercury in fish potentially alter androgen profiles, they do not substantially decrease other indicators of general and reproductive health.


Biphasic Regulation of Mesenchymal Genes Controls Fate Switches During Hematopoietic Differentiation of Human Pluripotent Stem Cells.
Epithelial-mesenchymal transition (EMT) or its reverse process mesenchymal-epithelial transition (MET) occurs in multiple physiological and pathological processes. However, whether an entire EMT-MET process exists and the potential function during human hematopoiesis remain largely elusive. Utilizing human pluripotent stem cell (hPSC)-based systems, it is discovered that while EMT occurs at the onset of human hematopoietic differentiation, MET is not detected subsequently during differentiation. Instead, a biphasic activation of mesenchymal genes during hematopoietic differentiation of hPSCs is observed. The expression of mesenchymal genes is upregulated during the fate switch from pluripotency to the mesoderm, sustained at the hemogenic endothelium (HE) stage, and attenuated during hemogenic endothelial cell (HEP) differentiation to hematopoietic progenitor cells (HPCs). A similar expression pattern of mesenchymal genes is also observed during human and murine hematopoietic development in vivo. Wnt signaling and its downstream gene SNAI1 mediate the up-regulation of mesenchymal genes and initiation of mesoderm induction from pluripotency. Inhibition of transforming growth factor-β (TGF-β) signaling and downregulation of HAND1, a downstream gene of TGF-β, are required for the downregulation of mesenchymal genes and the capacity of HEPs to generate HPCs. These results suggest that the biphasic regulation of mesenchymal genes is an essential mechanism during human hematopoiesis.


Incidence and predictors of congestive heart failure after autologous hematopoietic cell transplantation.
Advances in autologous hematopoietic cell transplantation (HCT) strategies have resulted in a growing number of long-term survivors. However, these survivors are at increased risk of developing cardiovascular complications due to pre-HCT therapeutic exposures and conditioning and post-HCT comorbidities. We examined the incidence and predictors of congestive heart failure (CHF) in 1244 patients undergoing autologous HCT for a hematologic malignancy between 1988 and 2002. The cumulative incidence of CHF was 4.8% at 5 years and increased to 9.1% at 15 years after transplantation; the CI for female lymphoma survivors was 14.5% at 15 years. The cohort was at a 4.5-fold increased risk of CHF (standardized incidence ratio = 4.5), compared with the general population. The risk of CHF increased substantially for patients receiving ≥ 250 mg/m(2) of cumulative anthracycline exposure (odds ratio [OR]: 9.9, P < .01), creating a new and lower threshold for cardiac surveillance after HCT. The presence of hypertension among recipients of high-dose anthracycline (≥ 250 mg/m(2)) resulted in a 35-fold risk (OR: 35.3, P < .01) of CHF; the risk was nearly 27-fold (OR: 26.8, P < .01) for high-dose anthracycline recipients with diabetes, providing evidence that hypertension and diabetes may be critical modifiers of anthracycline-related myocardial injury after HCT and creating targeted populations for aggressive intervention.


1470 nm Diode Laser Enucleation vs Plasmakinetic Resection of the Prostate for Benign Prostatic Hyperplasia: A Randomized Study.
The purpose of the current work was to comparatively assess 1470 nm diode laser enucleation of the prostate (DiLEP) and plasmakinetic resection of the prostate (PKRP) for treating benign prostatic hyperplasia (BPH). From January 2016 to March 2017, 157 individuals with bladder outflow obstruction caused by BPH were randomized to DiLEP and PKRP groups, for prospective analysis. Of these, 152 cases were evaluated before operation and at 3, 6, and 12 months postsurgery. Patient baseline properties, presurgery data, and postsurgical outcomes were comparatively assessed, as well as complications. There were no significant preoperative differences between surgical groups. DiLEP-treated cases showed remarkable reduced operative time, postsurgical bladder irrigation time, catheterization duration, and hospital stay compared with the PKRP group (P < 0.001). Hemoglobin amount decrease was markedly less pronounced after DiLEP (P = 0.004). However, no patients needed blood transfusion in either group. The decrease in sodium level showed no marked differences between the DiLEP and PKRP groups (P = 0.380). In addition, complications were comparable and no significant differences in both groups. At 3, 6, and 12 months, International Prostate Symptom Score (IPSS), quality of life (QoL), maximum flow rate (Qmax), and postvoid residual (PVR) were similar in both groups (P > 0.05). DiLEP and PKRP are similar in efficacy and safety for relieving obstruction and low urinary tract symptoms. Compared with PKRP, DiLEP has decreased risk of hemorrhage, operative time, bladder irrigation time, catheterization duration, and hospital stay. However, IPSS, QoL, Qmax, and PVR were similar for both procedures within 12 postoperative months.


In vivo effect of human granulocyte colony-stimulating factor derivatives on hematopoiesis in primates.
Using cynomolgus monkeys, we studied the hematopoietic effect and antigenicity of the human granulocyte colony-stimulating factor (hG-CSF) derivatives mutated at N-terminal amino acids (NC-59 and KW-2228) and that lacking N-terminal amino acids (M-7) in comparison with intact hG-CSF. These compounds were subcutaneously administered daily into the back of cynomolgus monkeys at the doses of 1 and 100 micrograms/kg for 2 weeks, and withdrawn for 2 weeks (1st course). After that, the same schedule was repeated (2nd course), and intact hG-CSF and KW-2228 were further administered for 2 weeks (3rd course). In the first course, all of the G-CSF derivatives showed excellent dose-dependent granulopoietic activities. In the 2nd course, however, the activities of NC-59 and M-7 were decreased and the elevations of binding and neutralizing antibody titers were observed. On the other hand, KW-2228 showed granulopoietic activities equal to or better than that of intact hG-CSF and had low antibody titer throughout all the courses.


Facilitating Redox Cycles of Copper Species by Pollutants in Peroxymonosulfate Activation.
The redox behavior of metal active sites determines the rate of heterogeneous catalysis in peroxymonosulfate activation. Previous reports focused on the construction of catalysts for accelerating interfacial electron transfer. In this work, a new strategy was proposed for facilitating valence cycles of Cu+/Cu2+ by using pollutants. The 2.5Cu/CeO2/PMS system was capable of achieving the efficient removal of pollutants, including tetracycline, oxytetracycline, and rhodamine B, in a wide pH working range. In the presence of tetracycline, a Cu-N bond was formed between the -NH2 group of tetracycline and the Cu site of the catalyst, showing that the coordination of Cu active sites changed to CuO4N1. The charge of CuO4N1 active sites rearranged, making it easier to obtain electrons and promote the PMS oxidation, thereby accelerating the reduction of Cu2+ to Cu+ and PMS activation. The PMS activation system showed excellent sustainability and selectivity for the removal of organic pollutants. This study provides a novel routine to promote peroxymonosulfate activation by utilizing pollutants to accelerate the redox behavior of metal species.


Immunonegative Staining: Epitope Localization on Macromolecules
Relevant literature relating to immunonegative staining is reviewed and integrated with current research of the author and others. The immunonegative staining procedure has been utilized for the study of epitope localization on immune complexes formed from keyhole limpet hemocyanin type 2 (KLH2) di- and multidecamers, and the 20S and 26S proteasome from Xenopus laevis. The IgG linkage pattern of molecules in small immune complexes is considered to provide the most reliable indication of epitope location. For both KLH2 and the 20S proteasome, using domain-specific monoclonal antibodies and a 32-kDa (p32) subunit-specific polyclonal antibody, respectively, it is shown that epitopes (KLH2, subunit fg domain pair and the proteasome p32) are located on the ends of the two cylindrical molecules. Also, for KLH2 with a monoclonal antibody directed against subunit domain a, it is shown that this domain is located at the center of the KLH2 didecamer. Interpretation of the data for KLH2 indicates that the 10 subunits in each decamer are likely to be oriented in parallel. The epitopes of the 10 fg domain pairs are located on the ends, near the edge of the didecamer cylinder (i.e., at the "collar" region of the didecamer) and the epitope of the 10 a domains is located on the side wall of the didecamer cylinder, where the two decamers meet. In view of the varying shape of the Xenopus 20S proteasome immune complexes, it is tentatively proposed that there may be two p32 subunits located in each of the heptameric alpha rings rather than one, but the possibility of macromolecular heterogeneity with respect to p32 subunit composition cannot be discounted.


Aspartate aminotransferase in alfalfa root nodules : I. Purification and partial characterization.
Aspartate aminotransferase (l-aspartate:2-oxoglutarate aminotransferase, EC 126.96.36.199 [AAT]), a key enzyme in the assimilation of C and N compounds, was purified from the cytosol of alfalfa (Medicago sativa L.) root nodules. Isoforms that increased during nodule development, AAT-2a, AAT-2b, and AAT-2c, were purified greater than 447-fold to apparent homogeneity, and high titer polyclonal antibodies were produced. The native molecular weight of the AAT-2 isoforms was approximately 80 kilodatons with a subunit molecular weight of 40 kilodatons, indicating that the holoenzymes are dimers. The AAT-2 isoforms comprised approximately 0.4% of the total soluble nodule protein. The AAT specific activity was measured in leaf, stem, root, and nodule organs, and zymograms of each were compared. Enzyme activity was 4- to 37-fold greater in effective (nitrogen fixing) nodules than in leaves, stems, and roots. Effective nodule AAT-specific activity was 3- to 8-fold greater than that of plant-controlled ineffective nodules. No differences in K(m) were observed between AAT-1 and AAT-2. Antibodies raised against AAT-2 were more selective against AAT-2 than AAT-1. Evidence obtained from zymograms suggests that the expression of alfalfa nodule AAT is controlled at two different gene loci, AAT-1 and AAT-2, resulting in different dimeric isoforms.


γδ T cell migration: Separating trafficking from surveillance behaviors at barrier surfaces.
γδ T cells are found in highest numbers at barrier surfaces throughout the body, including the skin, intestine, lung, gingiva, and uterus. Under homeostatic conditions, γδ T cells provide immune surveillance of the epidermis, intestinal, and oral mucosa, whereas the presence of pathogenic microorganisms in the dermis or lungs elicits a robust γδ17 response to clear the infection. Although T cell migration is most frequently defined in the context of trafficking, analysis of specific migratory behaviors of lymphocytes within the tissue microenvironment can provide valuable insight into their function. Intravital imaging and computational analyses have been used to define "search" behavior associated with conventional αβ T cells; however, based on the known role of γδ T cells as immune sentinels at barrier surfaces and their TCR-independent functions, we put forth the need to classify distinct migratory patterns that reflect the surveillance capacity of these unconventional lymphocytes. This review will focus on how γδ T cells traffic to various barrier surfaces and how recent investigation into their migratory behavior has provided unique insight into the contribution of γδ T cells to barrier immunity.


Giant extended pi-conjugated dendrimers containing the 10,15-dihydro-5H-diindeno[1,2-a;1',2'-c]fluorene chromophore: synthesis, NMR behaviors, optical properties, and electroluminescence.
This paper reports the facile synthetic strategy of a series of novel pi-conjugated dendrimers (G0 and G1) based on 10,15-dihydro-5H-diindeno[1,2-a;1',2'-c]fluorene (truxene) in which the benzene cores are generated "in-situ" from acetyl aromatics by the acid-promoted cyclotrimerizations. The unique NMR behaviors, physical properties, and electroluminescence device applications are also presented. Besides the purity, the structure, and the confirmation of the successful formation and isolation of desired compounds by clear assignments of every molecule by (1)H and (13)C and 2D NMR characterizations, several astonishing NMR behaviors have been observed in various solvents. For 1, chemical shift values belonging to H-2' of hexyl substituents move to the most upfield; however, such chemical shift values move from 0.48 to 0.85 ppm when pyridine-d(5) or benzene-d(6) is used as solvent. Our dendrimers as the emissive layers in organic light-emitting diodes gave blue-green light with an external quantum efficiency up to 0.16% both for G0 and for G1 in nitrogen, respectively, which exhibit unique electroluminescence spectra in comparison with their corresponding photoluminescence ones.


Effects of PN 200-110, a new calcium antagonist, on alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction elicited by selective alpha-adrenoceptor agonists and catecholamines in the pithed rat.
The inhibitory action of PN 200-110, a novel calcium antagonist, on alpha 1- and alpha 2-adrenoceptor mediated pressor effects elicited by selective alpha-adrenoceptor agonists and catecholamines was studied in pithed rats. The pressor effects of the selective alpha 1-adrenoceptor agonist cirazoline were only slightly influenced by PN 200-110 (0.1 mg/kg). On the other hand, PN 200-110 efficiently inhibited the hypertensive effects of the selective alpha 2-adrenoceptor agonist B-HT 920 in a dose-dependent way, thereby reducing the maximal response considerably (pD2' = 7.30). The alpha 1-adrenoceptor mediated pressor responses of adrenaline and noradrenaline, obtained after pretreatment with propranolol and the selective alpha 2-adrenoceptor antagonist yohimbine, were slightly inhibited by PN 200-110, without influencing maximal responses. PN 200-110 inhibited the alpha 1-adrenergic pressor effects of noradrenaline better than those of adrenaline. The alpha 2-adrenoceptor mediated pressor effects of the cathecholamines obtained after pretreatment with propranolol and the selective alpha 1-adrenoceptor antagonist prazosin, were highly susceptible to blockade by PN 200-110 except for the high doses (300 micrograms/kg) of adrenaline. The present study establishes the pronounced calcium entry inhibitory potency of PN 200-110 and confirms and extends previous observations that blockade of calcium entry preferably impairs alpha 2-adrenoceptor mediated vasoconstriction in vivo, when compared to that elicited by alpha 1-adrenoceptor stimulation.


Lobular Mammary Carcinoma Presenting as an Obstructing Rectal Mass.
Breast cancer is the leading cause of cancer death in women, and while metastasis is common to areas like the bone, lungs, and brain, it is rare to metastasize to the gastrointestinal tract and especially to the rectum. Due to the rarity of this condition and its resemblance clinically and radiologically to primary gastrointestinal tract tumors, diagnosis and treatment are challenging. We present a case of metastatic lobular mammary carcinoma in a 52-year-old Bahraini woman who presented with an obstructing rectal mass.


Gastrointestinal and hepatic side effects of potential treatment for COVID-19 and vaccination in patients with chronic liver diseases.
The outbreak of coronavirus disease 2019 (COVID-19) is a global pandemic. Many clinical trials have been performed to investigate potential treatments or vaccines for this disease to reduce the high morbidity and mortality. The drugs of higher interest include umifenovir, bromhexine, remdesivir, lopinavir/ritonavir, steroid, tocilizumab, interferon alpha or beta, ribavirin, fivapiravir, nitazoxanide, ivermectin, molnupiravir, hydroxychloroquine/chloroquine alone or in combination with azithromycin, and baricitinib. Gastrointestinal (GI) symptoms and liver dysfunction are frequently seen in patients with COVID-19, which can make it difficult to differentiate disease manifestations from treatment adverse effects. GI symptoms of COVID-19 include anorexia, dyspepsia, nausea, vomiting, diarrhea and abdominal pain. Liver injury can be a result of systemic inflammation or cytokine storm, or due to the adverse drug effects in patients who have been receiving different treatments. Regular monitoring of liver function should be performed. COVID-19 vaccines have been rapidly developed with different technologies including mRNA, viral vectors, inactivated viruses, recombinant DNA, protein subunits and live attenuated viruses. Patients with chronic liver disease or inflammatory bowel disease and liver transplant recipients are encouraged to receive vaccination as the benefits outweigh the risks. Vaccination against COVID-19 is also recommended to family members and healthcare professionals caring for these patients to reduce exposure to the severe acute respiratory syndrome coronavirus 2 virus.


From vineyard to winery: a source map of microbial diversity driving wine fermentation.
Humans have been making wine for thousands of years and microorganisms play an integral part in this process as they not only drive fermentation, but also significantly influence the flavour, aroma and quality of finished wines. Since fruits are ephemeral, they cannot comprise a permanent microbial habitat; thus, an age-old unanswered question concerns the origin of fruit and ferment associated microbes. Here we use next-generation sequencing approaches to examine and quantify the roles of native forest, vineyard soil, bark and fruit habitats as sources of fungal diversity in ferments. We show that microbial communities in harvested juice and ferments vary significantly across regions, and that while vineyard fungi account for ∼40% of the source of this diversity, uncultivated ecosystems outside of vineyards also prove a significant source. We also show that while communities in harvested juice resemble those found on grapes, these increasingly resemble fungi present on vine bark as the ferment proceeds.


Four new cases of lethal osteogenesis imperfecta due to glycine substitutions in COL1A1 and genes. Mutations in brief no. 152. Online.
Perinatal lethal osteogenesis imperfecta is the result of heterozygous mutations of the COL1A1 and COL1A2 genes. Here we describe the molecular defects responsible for four case of lethal OI. Two glycine substitutions within the COL1A1 gene (G478S, G994D) and two glycine substitutions within the COLIA2 gene (G319V, G697C) were identified. The mutation sites were localized in proalpha2(I) and proalpha2(I)mRNA molecules, respectively, by chemical cleavage of mismatch in hereteroduplex nucleic acids. Subsequent reverse transcription PCR amplification, cloning and sequencing, led to mutation identification. The aminoacid substitutions were due to two G-->A transitions in COL1A1(cases 1,2), to a G-->T transversion in COL1A2 (case 3), and to two contiguous point mutations in COL1A2 (case 4). All five nucleotide changes appeared to be fresh mutations. COLIA1(accession number Z74615) and COL1A2(accession number Z74616) wild type coding sequences (cDNA) were deduced from the EMBL DNA sequence database. The mutations described here can also be found in the human type I collagen mutation database at the web site:http://www.le.ac.uk/genetics/collagen.


Role of tissue and circulating microRNAs and DNA as biomarkers in medullary thyroid cancer.
Medullary thyroid carcinoma (MTC) is a rare neuroendocrine tumor comprising hereditary or sporadic form with frequent mutations in the rearranged during transfection (RET) or RAS genes. Diagnosis is based on the presence of thyroid tumor mass with altered levels of calcitonin (Ctn) and carcinoembryonal antigen (CEA) in the serum and/or in the cytological smears from fine needle aspiration biopsies. Treatment consists of total thyroidectomy, followed by tyrosine kinase inhibitors (TKi) in case of disease persistence. During TKi treatment, Ctn and CEA levels can fluctuate regardless of tumor volume, metastasis or response to therapy. Research for more reliable non-invasive biomarkers in MTC is still underway. In this context, circulating nucleic acids, namely circulating microRNAs (miRNAs) and cell free DNA (cfDNA), have been evaluated by different research groups. Aiming to shed light on whether miRNAs and cfDNA are suitable as MTC biomarkers we searched three different databases, PubMed, Scopus, WOS and reviewed the literature. We classified 83 publications fulfilling our search criteria and summarized the results. We report data on miRNAs and cfDNA that can be evaluated for validation in independent studies and clinical application.


The expression patterns of a eukaryotic initiation factor 3 subunit H in the silk glands in Bombyx mori.
Eukaryotic initiation factor 3 subunit H has been characterized in many organisms, and it has been found to play many roles including help regulate translation initiation. In this work, we studied the tissue distribution and expression profiles of Bombyx mori (B. mori) eIF3 subunit H (BmeIF3h). BmeIF3h was prominently expressed in silk glands, with anterior silk glands (ASGs), middle silk glands (MSGs), and posterior silk glands (PSGs) all expressing BmeIF3h. The expression levels of BmeIF3h in MSGs and PSGs were higher than that in ASGs during 0 d and 2 d of the 5th instar larvae. The expression levels of BmeIF3h in MSGs and PSGs were up-regulated once the silk glands began to synthesize silk protein during the feeding stage of the 4th instar larvae. Immunohistochemistry showed that BmeIF3h was distributed in the cytoplasm of MSGs cells and in both the nucleus and the cytoplasm of PSGs cells. These data suggest that BmeIF3h had different action behaviors in the MSGs and PSGs related to the production of the silk glue proteins and silk fibre proteins, respectively.


The Intricate Crosstalk Between Insulin and Pancreatic Ductal Adenocarcinoma: A Review From Clinical to Molecular.
Increased insulin level (or "hyperinsulinemia") is a common phenomenon in pancreatic ductal adenocarcinoma (PDA) patients and signals poor clinical outcomes. Insulin is safe in low PDA risk population, while insulin significantly promotes PDA risk in high PDA risk population. The correlation between insulin and PDA is a reciprocal self-reinforcing relationship. On the one hand, pancreatic cancer cells synthesize multiple molecules to cause elevated peripheral insulin resistance, thus enhancing hyperinsulinemia. On the other hand, insulin promotes pancreatic cancer initiation and sustains PDA development by eliciting tumorigenic inflammation, regulating lipid and glucose metabolic reprogram, overcoming apoptosis through the crosstalk with IGF-1, stimulating cancer metastasis, and activating tumor microenvironment formation (inflammation, fibrosis, and angiogenesis). Currently, taking glucose sensitizing agents, including metformin, SGLT-2 inhibitor, and GLP-1 agonist, is an effective way of lowering insulin levels and controlling PDA development at the same time. In the future, new drugs targeting insulin-related signal pathways may pave a novel way for suppressing PDA initiation and progression.


Co-transport of Potassium and Sugars across the Plasmalemma of Mesophyll Protoplasts.
Sugars (sucrose + hexoses) produced photosynthetically by isolated mesophyll protoplasts of wheat and tobacco were effluxed across the plasma membrane (3 to 10 micromoles hexose equivalents per milligram chlorophyll per hour). The efflux was sensitive to uncouplers and oligomycin which indicated a requirement for energy. A proton gradient was probably not coupled directly to the transport because changing the proton gradient across the plasma membrane by varying the pH of the medium or by adding sodium acetate had no significant effect on the rate of sugar release.A release of K(+) was associated with sugar efflux from the protoplasts. The molar ratio of K(+) to sugar varied between 1.5 and 2.5, depending on the species. Exogenous CKl, RbCl, and LiCl (50 millimolar each), but not NaCl or CsCl, significantly inhibited sugar efflux. Conditions that reduced sugar efflux (exogenous KCl, LiCl, mersalyl, or oligomycin) also reduced K(+) release and caused a time-dependent reduction in photosynthetic sucrose formation and increased amino acid and starch formation. Results obtained support the postulate that a K(+) symport is involved in the transport of sugar across the energized plasmalemma of photosynthetically active mesophyll cells.


Importance of assisted reproductive technologies in the conservation of wild, rare or indigenous ungulates: review article.
Biodiversity is increasingly threatened by intensive agriculture, environmental pollution, extinction of natural habitats and several other factors. Several mammalian species including ungulates have disappeared or are threatened by extinction. However, ungulates play an important role both in the ecosystem and in the economy. In general, species or breeds are considered endangered if their population does not exceed 1,000 individuals. In these cases conservation programmes should be initiated in order to maintain or even increase their number. This review deals with the possibilities and limitations of assisted reproductive technologies (ART) in the conservation of ecologically valuable wild, rare and indigenous ungulates. The methods discussed here are artificial insemination, cryopreservation of semen and embryos, embryo recovery and transfer, in vitro production of embryos, as well as micromanipulation techniques including sperm injection, assisted hatching and cloning. Some of these procedures are already being exploited in the breeding of farm ungulates, but more basic information about the reproductive patterns of wild, rare and indigenous animal species is needed before the routine use of ARTs.


Expression and splicing of ABC and SLC transporters in the human blood-brain barrier measured with RNAseq.
The blood-brain barrier (BBB) expresses numerous membrane transporters that supply needed nutrients to the central nervous system (CNS), consisting mostly of solute carriers (SLC transporters), or remove unwanted substrates via extrusion pumps through the action of ATP binding cassette (ABC) transporters. Previous work has identified many BBB transporters using hybridization arrays or qRT-PCR, using targeted probes. Here we have performed next-generation sequencing of the transcriptome (RNAseq) extracted from cerebral cortex tissues and brain microvessel endothelial cells (BMEC) obtained from two donors. The same RNA samples had previously been measured for transporter expression using qRT-PCR (Geier et al., 2013), yielding similar expression levels for overlapping mRNAs (R=0.66, p<0.001). RNAseq confirms a number of transporters highly enriched in BMECs (e.g., ABCB1, ABCG2, SLCO2B1, and SLC47A1), but also detects novel BMEC transporters. Multiple splice isoforms detected by RNAseq are either robustly enriched or depleted in BMECs, indicating differential RNA processing in the BBB. The Complete RNAseq data are publically available (GSE94064).


Effects of neutral phytase on growth performance and phosphorus utilization in crucian carp (Carassius auratus).
A feeding trial was conducted for nine weeks to investigate the effects of partially replacing Ca(H2PO4)2 with neutral phytase on the growth performance, phosphorus utilization, nutrient digestibility, serum biochemical parameters, bone and carcass mineral composition, and digestive-enzyme-specific activity in crucian carp (Carassius auratus). The diets prepared with 0.8%, 0%, and 1.8% Ca(H2PO4)2 (1%=1 g/100 g) supplements were regarded as the P1E0, negative control (NC), and positive control (PC) groups, respectively; the other three experimental diets were prepared with the addition of 200, 300, and 500 U/kg of neutral phytase, respectively, based on the P1E0 group. Three hundred and eighty-four fish ((1.50±0.01) g) were randomly distributed in the six treatments with four replicates each. The fish were initially fed with 2%-3% diets of their body weight per day, with feeding twice daily (08:00 and 16:00), under a 12-h light/12-h dark cycle at the temperature of (27.56±0.89) °C. The results showed that supplemental phytase at different levels in the diet improved the final body weight, average daily gain, feed conversion ratio, phosphorus utilization, and protein efficiency ratio of crucian carp (P<0.05). Phytase supplementation increased the mineral content in serum (P), bone (P, Ca), and carcass (P, Ca, Zn, Na, and Mg) (P<0.05); the trypsin and chymotrypsin activity soared when fed with the phytase-supplemented diets (P<0.05). We may conclude that supplemental dietary neutral phytase improved the growth performance, phosphorus utilization as well as nutrient utilization in crucian carp, and it can be considered an important nutritional replacement for Ca(H2PO4)2.


An assessment of the plastic Thermafil obturation technique. Part 2. Material adaptation and sealability.
The short- and long-term apical seal of root canals obturated with softened alpha-phase gutta-percha on plastic core-carriers, Thermafil, was compared to that of laterally condensed, cold gutta-percha. Fifty-one roots from mandibular molars with separate canals, patent canal orifices and curvatures greater than 15 degrees were cleaned and shaped with K-files and 2.5% sodium hypochlorite to a size 30 at the apex, and flared with Hedstrom files to create a continuously tapering funnel preparation. Canals were randomly obturated with Sealapex root canal sealer and either alpha-phase gutta-percha on a plastic Thermafil carrier, or standard beta-phase gutta-percha with lateral condensation. Teeth were separated into three groups of 17 each and placed in black India ink for 24 h, 7 days or after 5 months storage in water. The teeth were demineralized, rendered transparent, and apical microleakage determined by the linear measurement of dye penetration. Significant differences in microleakage were noted between the 24 h and 5-month Thermafil groups (P < 0.05), 24-h and 7-day lateral condensation groups (P < 0.05), and 24-h and 5-month lateral condensation groups (P < 0.05). There were no significant differences between the two techniques at each time interval. It was concluded that both techniques demonstrated a significant increase in apical microleakage over a 5-month period.


LINC02875 Upregulation Contributed to Poor Prognosis for the Hepatocellular Carcinoma and Progression for the Cancerous Cells.
The prognostic implications and physiological effect of LINC02875 are unknown in hepatocellular carcinoma (HCC). We sought to examine the prognostic value of LINC02875 in HCC and assessed its role in HCC cellular function. LINC02875 expression was evaluated by RT-qPCR in HCC specimens and cell lines. LINC02875 expression was subjected to assess the correlation with clinical parameters by Chi-squared test and overall survival by Kaplan - Meier curve and Cox regression analysis. The effects of LINC02875 on the biological characteristics of HCC cells were studied by MTS and Transwell assay. LINC02875 was high-expressed in HCC, and this was associated with unfavorable clinical features and poor prognosis of HCC, especially HBV-related HCC. Knockdown of LINC02875 inhibited the proliferation, migration, and invasion of HCC cells. miR-485-5p was a downstream microRNA of LINC02875. LINC02875 affects the prognosis of HCC patients, especially HBV-related ones. LINC02875 represents a suitable therapeutic target for HCC.


Gut-Restricted Selective Cyclooxygenase-2 (COX-2) Inhibitors for Chemoprevention of Colorectal Cancer.
Selective cyclooxygenase (COX)-2 inhibitors have been extensively studied for colorectal cancer (CRC) chemoprevention. Celecoxib has been reported to reduce the incidence of colorectal adenomas and CRC but is also associated with an increased risk of cardiovascular events. Here, we report a series of gut-restricted, selective COX-2 inhibitors characterized by high colonic exposure and minimized systemic exposure. By establishing acute ex vivo 18F-FDG uptake attenuation as an efficacy proxy, we identified a subset of analogues that demonstrated statistically significant in vivo dose-dependent inhibition of adenoma progression and survival extension in an APCmin/+ mouse model. However, in vitro-in vivo correlation analysis showed their chemoprotective effects were driven by residual systemic COX-2 inhibition, rationalizing their less than expected efficacies and highlighting the challenges associated with COX-2-mediated CRC disease chemoprevention.


Catalytic roles for carbon-oxygen hydrogen bonding in SET domain lysine methyltransferases.
SET domain enzymes represent a distinct family of protein lysine methyltransferases in eukaryotes. Recent studies have yielded significant insights into the structural basis of substrate recognition and the product specificities of these enzymes. However, the mechanism by which SET domain methyltransferases catalyze the transfer of the methyl group from S-adenosyl-L-methionine to the lysine epsilon-amine has remained unresolved. To elucidate this mechanism, we have determined the structures of the plant SET domain enzyme, pea ribulose-1,5 bisphosphate carboxylase/oxygenase large subunit methyltransferase, bound to S-adenosyl-L-methionine, and its non-reactive analogs Aza-adenosyl-L-methionine and Sinefungin, and characterized the binding of these ligands to a homolog of the enzyme. The structural and biochemical data collectively reveal that S-adenosyl-L-methionine is selectively recognized through carbon-oxygen hydrogen bonds between the cofactor's methyl group and an array of structurally conserved oxygens that comprise the methyl transfer pore in the active site. Furthermore, the structure of the enzyme co-crystallized with the product epsilon-N-trimethyllysine reveals a trigonal array of carbon-oxygen interactions between the epsilon-ammonium methyl groups and the oxygens in the pore. Taken together, these results establish a central role for carbon-oxygen hydrogen bonding in aligning the cofactor's methyl group for transfer to the lysine epsilon-amine and in coordinating the methyl groups after transfer to facilitate multiple rounds of lysine methylation.


Sensory loss in syringomyelia: not necessarily dissociated.
The pattern of sensory loss was assessed in 210 cases of syringomyelia. Dissociated sensory loss occurred in 49% of cases, indicating that its occurrence is not a necessary finding for the diagnosis of syringomyelia. Syringomyelia should be considered in the differential diagnosis of all cases of spinal cord disease.


Investigation of reactive properties, adsorption on fullerene, DFT, molecular dynamics simulation of an anthracene derivative targeting dihydrofolate reductase and human dUTPase.
Anthracenes are aromatic compounds with flexible structure and reactivity which are of great interest to theoretical and experimental chemists. Theoretical investigations of 1,4-dihydroxy-5,8-bis[2-(2-hydroxyethylamino)ethylamino]anthracene-9,10-dione (Mitoxantrone) (DDEA) based on density functional theory, molecular dynamics and adsorption on fullerene are reported in the present research. The suitable situation for adsorption with fullerene (C60) is the cyclohex-2-ene-1,4-dione ring of DDEA. Selected quantum-molecular descriptors have been calculated to predict the most reactive sites of the DDEA molecule. Interactions of DDEA with water have been studied using MD simulations. MD simulations were also used to study solubility parameter, a significant quantity for the development of pharmaceutical formulations. The affinity of DDEA on human dihydrofolate reductase and deoxyuridine triphosphatase enzymes was investigated by MD simulation of the protein-ligand complex obtained by molecular docking study.Communicated by Ramaswamy H. Sarma.


Serologic and Molecular Evidence of Widespread Infection of Avian Hepatitis E Virus in Poultry Farms of Iran.
Hepatitis-splenomegaly syndrome is caused by avian hepatitis E virus (aHEV), a nonenveloped, single-stranded RNA virus. The economic importance of this disease in the poultry industry is due to the decline in egg production (10%-40%) and the rise in mortality (1%-4%). In the present study, 1540 serum samples from 33 broiler breeder flocks were analyzed by an enzyme-linked immunosorbent assay for the presence of an anti-aHEV antibody. In addition, a diagnostic nested reverse transcriptase-PCR was done on all farm samples. In the serologic study, 66.7% (22/33) of the flocks and 28.5% (439/1540) of the chickens were positive. The molecular study showed that three farms were positive, and PCR products were observed for the conserved regions of the aHEV helicase and capsid virus genes as 386 bp and 242 bp, respectively. It should be noted that clinical and pathologic symptoms including decreased egg production, enlarged livers and spleens, and a slight rise in mortality rate were observed in eight farms. To our knowledge, this is the first documented study on the aHEV identification and its antibody detection in broiler breeder farms in Iran.


Retrobulbar optic neuritis and rhegmatogenous retinal detachment in a fourteen-year-old girl with retinitis pigmentosa sine pigmento.
A 14-year-old girl complained of a sudden decrease in right visual acuity. The patient had night blindness, a mottled retina but no pigments, extinguished scotopic electroretinographic response, central scotoma in the right eye and rhegmatogenous retinal detachment. She had initially received laser photocoagulation around the retinal tear and then corticosteroid therapy, cryoretinopexy and segmental buckling. Her right visual acuity increased to 1.0. The association of retinitis pigmentosa sine pigmento, retrobulbar optic neuritis and rhegmatogenous retinal detachment, as demonstrated in our patient, may be uncommon.


Factors affecting ice nucleation in plant tissues.
Factors affecting the ice nucleation temperature of plants and plant tissues were examined. The mass of a sample had a marked effect on ice nucleation temperature. Small tissue samples supercooled to -10 degrees C and were not accurate predictors of the nucleation temperature of intact plants in either laboratory or field experiments. This effect was not unique to plant tissues and was observed in autoclaved and control soil samples. Ice nucleation temperatures of bean, corn, cotton, and soybean seedlings were influenced by the length of subzero exposure, presence of ice nucleation active bacteria, and leaf surface wetness. The number of factors influencing ice nucleation temperature suggested that predicting the freezing behavior of plants in the field will be complex.


In vitro analysis of the role of glucose oxidase from Talaromyces flavus in biocontrol of the plant pathogen Verticillium dahliae.
Culture filtrates from Talaromyces flavus grown on glucose contained high levels of glucose oxidase activity, while culture filtrates from T. flavus grown on xylan contained negligible glucose oxidase activity. Culture filtrates from T-flavus grown on both media contained complex protein profiles. However, only culture filtrates from T. flavus grown on glucose inhibited germination of microsclerotia of Verticillium dahliae in in vitro inhibition assays. A polyclonal antiserum preparation, pABGO-1, raised against purified glucose oxidase from T. flavus was highly specific for glucose oxidase. Only one protein band in culture filtrates (from glucose medium), migrating at 71 kDa, was detected in Western blots (immunoblots) with this antiserum. This band comigrated with purified glucose oxidase. No bands were detected in culture filtrates from the xylan medium. Glucose oxidase was removed via immunoprecipitation from culture filtrates of T. flavus grown in glucose medium, resulting in filtrates which no longer inhibited in vitro microsclerotial germination. When glucose oxidase-depleted filtrates were amended with purified glucose oxidase from T. flavus, the ability to kill microsclerotia in vitro was restored to original levels. We conclude that glucose oxidase is the only protein in culture filtrates of T. flavus responsible for inhibition of germination of microsclerotia of V. dahliae.


Elective superior mediastinal dissection for laryngeal carcinoma involving subglottis.
Despite the low incidence of superior mediastinal metastasis (SMM) we recommend elective superior mediastinal dissection (ESMD) and close follow-up for all patients with laryngeal carcinoma involving the subglottis, because higher peristomal recurrence (PR) rates and lower survival rates were observed in the presence of SMM numerically even though statistical support was lacking. This study was undertaken to identify the incidence of SMM and to evaluate the need for ESMD in laryngeal tumors involving the subglottis. Medical records of patients who had undergone total laryngectomy for laryngeal carcinoma were reviewed retrospectively and 68 patients (45 with ESMD and 23 without ESMD) who had pathologically proven subglottic involvement were included in the study. SMM was found in 3 of 45 (6.6%) patients who underwent ESMD. The PR rate was higher in patients with SMM (66.7%) than in patients without SMM (9.5%) and the presence of SMM was significantly related to PR (p = 0.043) in univariate analysis. However, this relationship was not preserved in multivariate analysis. The 3- and 5-year survival rates of patients with SMM were lower (both 66.7%) than patients without SMM (83.3% and 76.2%, respectively), but decreased survival rates did not differ statistically from the survival rates of 42 patients without SMM.


Focusing single-order diffraction transmission grating with a focusing plane perpendicular to the grating surface.
By combining the single-order dispersion properties of quasi-sinusoidal single-order diffraction transmission gratings (QSTG) and the single-foci focusing properties of annulus-sector-shaped-element binary Gabor zone plate (ASZP), we propose a novel focusing single-order diffraction transmission grating (FSDTG). Different from the diffraction patterns of a normal transmission grating (TG), it has a focusing plane perpendicular to the grating surface. Numerical simulations are carried out to verify its diffraction patterns in the framework of Fresnel-Kirchhoff diffraction. Higher-order diffraction components of higher harmonics can be effectively suppressed by the FSDTG we designed. And we find that the focal depth and resolving power are only determined by the structure parameters of our FSDTG by theoretical estimations.


Engineered peptide-based nanobiomaterials for electrochemical cell chip.
Biomaterials having cell adhesion ability are considered to be integral part of a cell chip. A number of researches have been carried out to search for a suitable material for effective immobilization of cell on substrate. Engineered ECM materials or their components like collagen, Poly-l-Lysine (PLL), Arg-Gly-Asp (RGD) peptide have been extensively used for mammalian cell adhesion and proliferation with the aim of tissue regeneration or cell based sensing application. This review focuses on the various approaches for two- and three-dimensionally patterned nanostructures of a short peptide i.e. RGD peptide on chip surfaces together with their effects on cell behaviors and electrochemical measurements. Most of the study concluded with positive remarks on the well-oriented engineered RGD peptide over their homogenous thin film. The engineered RGD peptide not only influences cell adhesion, spreading and proliferation but also their periodic nano-arrays directly influence electrochemical measurements of the chips. The electrochemical signals found to be enhanced when RGD peptides were used in well-defined two-dimensional nano-arrays. The topographic alteration of three-dimensional structure of engineered RGD peptide was reported to be suitably contacted with the integrin receptors of cellular membrane which results indicated the enhanced cell-electrode adhesion and efficient electron exchange phenomenon. This enhanced electrochemical signal increases the sensitivity of the chip against the target analytes. Therefore, development of engineered cellular recognizable peptides and its 3D topological design for fabrication of cell chip will provide the synergetic effect on bio-affinity, sensitivity and accuracy for the in situ real-time monitoring of analytes.


Increases in myocardial cyclic GMP attenuate contractile delay in myocardial stunning.
We tested the hypothesis that the effects of myocardial stunning would be reduced by cyclic GMP in rabbit hearts. In three groups of anesthetized open-chest New Zealand white rabbits, myocardial stunning was produced by 15 min of occlusion of the left anterior descending coronary artery followed by 15 min of reperfusion repeated twice. Either control vehicle (saline plus 1% dimethyl sulfoxide) or 8-bromo-cyclic GMP (8-Br-cGMP (10(-4) and 10(-3) M)) was topically applied to the left ventricular surface. Hemodynamic (left ventricular and aortic pressures) and functional parameters (wall thickening, delay in onset of wall thickening, and rate of wall thickening) were determined. Coronary blood flow (microspheres) and O2 extraction (microspectrophotometry) were used to determine myocardial O2 consumption (VO2). Myocardial stunning was observed in the control group through an increased delay in onset of myocardial wall thickening (29 +/- 7 versus 55 +/- 16 ms) and decreased maximal rate of wall thickening (20 +/- 8 versus 11 +/- 3 mm x s(-1)). After treatment with 8-Br-cGMP 10(-4) and 10(-3) M, stunning did not increase the delay (37 +/- 5 versus 39 +/- 7 and 39 +/- 7 versus 28 +/- 8 ms). Myocardial stunning did not significantly alter VO2. 8-Br-cGMP 10(-3) M significantly decreased subepicardial VO2 (6.2 +/- 0.8 versus 3.7 +/- 0.6 mL O2 x min(-1) 100 g(-1)) and insignificantly decreased subendocardial VO2 (8.6 +/- 0.9 versus 6.3 +/- 1.2 mL O2 x min(-1) x 100 g(-1)) when compared with the vehicle-treated rabbits. We conclude that increasing cyclic GMP reduced the effects of myocardial stunning in the rabbit heart by ameliorating the delay in onset of wall thickening and decreasing the local O2 costs in the stunned region.


Effective orbital volume and eyeball position: an MRI study.
Previous studies have examined factors affecting the position of the eyeball to the orbit. This study examined the role of effective orbital volume (EOV), defined as the difference between orbital and eyeball volume, as a determinant of eyeball position, using MRI scans. Forty-six patients were recruited from the Department of Ophthalmology of the University Hospital of Heraklion, Crete Greece. Patients with a history of orbital disease were excluded. Distances between eyeball poles and orbital landmarks were measured in T1 weighted transverse, sagittal and coronal orbital images. The protrusion of the eyeball in the sagittal and transverse planes was recorded. The volume of the eyeball and bony orbit, the EOV, the volume of the extraocular muscles as well as clinical information (age, gender, Hertel exophthalmometry) were also recorded. EOV was significantly associated with orbital volume but not with eyeball volume. EOV was also significantly associated with transverse and sagittal globe protrusions. Females displayed significantly lower orbital and eyeball volumes as well as EOV than males but higher transverse globe protrusion than males. Variations in EOV are associated with orbital volume rather than with eyeball volume. EOV is associated with globe protrusion and may be taken into account in the planning of various procedures, including orbital decompression, treatment of enophthalmos or the size of orbital implants following enucleation.


Growing up with cerebral palsy: perceptions of the influence of family.
Cerebral palsy (CP) is a non-progressive condition present from birth or infancy that includes various neurological patterns of dysfunction. It is characterized by abnormal motor control and/or posture and can involve communication difficulties. Children and youth with CP face multiple social and developmental challenges during their formative years including mild to severe physical limitations, poor socialization, limited recreational activities, and stigmatization. Families play a key role in supporting adaptation to CP. The purpose of this paper was to explore women's perceptions of the ways their families contributed to their overall quality of life with CP. Drawing on data from a qualitative study of eight women with CP, the purpose of this analysis was to examine perceptions of the ways in which their families and individual family members contributed to participants' overall quality of life and adaptation to CP. Respondents ranged in age from 22 years to 55 years and had varied forms of CP. The study was based on a feminist biographical approach, which combines biographical methods with feminist principles. Participants provided considerable, rich contextual data on their family life and the pivotal role family played. The analysis identified four themes related to supportive family roles: (1) being an advocate, and teaching advocacy; (2) promoting inclusion and acceptance; (3) integrating therapy into daily life; and (4) the importance of siblings as friends and mentors. Health-care providers can contribute to the family's ability to facilitate quality of life by providing guidance on how to be advocates and teach advocacy, including the child with CP in family activities, accessing therapy and incorporating beneficial therapies at home, and promoting healthy sibling relationships.


Trichinella spiralis: Knockdown of gamma interferon inducible lysosomal thiol reductase (GILT) results in the reduction of worm burden.
Trichinella spiralis is mammalian skeletal muscles parasite which may cause trichinellosis in animals and humans. Gamma interferon inducible lysosomal thiol reductase (GILT) is a widespread superfamily which plays key role in processing and presentation of MHC class II restricted antigen by catalyzing disulfide bond reduction. There are no reports about GILT in T. spiralis. In present study, GILT from T. spiralis (Tsp-GILT) was cloned, analyzed by multiple-sequence alignment, and predicted by 3D structure model. Recombinant Tsp-GILT (about 46 kDa) was efficiently expressed in Escherichia coli and thiol reductase activity suggested that in acidic environment the addition of a reducing agent is needed. Soaking method was used to knockdown expression of Tsp-GILT using small interference RNA (siRNA). Immunofluorescence assay confirmed the transformation of siRNA into muscle larva (ML) and new born larva (NBL). Quantitative real time-PCR (QRT-PCR) analysis revealed that transcription level of Tsp-GILT mRNA can be up-regulated by stimulation of mouse IFN-γ and down-regulated by siRNA2 in vitro. NBLs soaked with siRNA2 showed 32.3% reduction in the generation of MLs. MLs soaked with siRNA2 showed 26.2% reduction in the next generation of MLs, but no significant effect was observed on adult worms or NBLs. These findings concluded that GILT may play important roles in the development of T. spiralis parasite.


Response of fluctuating and directional asymmetry to selection on wing shape in Drosophila melanogaster.
We tested whether directional selection on an index-based wing character in Drosophila melanogaster affected developmental stability and patterns of directional asymmetry. We selected for both an increase (up selection) and a decrease (down selection) of the index value on the left wing and compared patterns of fluctuating and directional asymmetry in the selection index and other wing traits across selection lines. Changes in fluctuating asymmetry across selection lines were predominantly small, but we observed a tendency for fluctuating asymmetry to decrease in the up-selected lines in both replicates. Because changes in fluctuating asymmetry depended on the direction of selection, and were not related to changes in trait size, these results fail to support existing hypotheses linking directional selection and developmental stability. Selection also produced a pattern of directional asymmetry that was similar in all selected lines whatever the direction of selection. This result may be interpreted as a release of genetic variance in directional asymmetry under selection.


Towards an anti-disease malaria vaccine.
Human infective parasites, such as those that cause malaria, are highly adapted to evade clearance by the immune system. In situations where they must maintain prolonged interactions with molecules of their host, they often use parasite surface protein families. These families are highly diverse to prevent immune recognition, and yet, to promote parasite survival, their members must retain the ability to interact with specific human receptors. One of the best understood of the parasite surface protein families is the PfEMP1 proteins of Plasmodium falciparum. These molecules cause infected erythrocytes to adhere to human receptors found on blood vessel and tissue surfaces. This protects the parasite within from clearance by the spleen and also causes symptoms of severe malaria. The PfEMP1 are exposed to the immune system during infection and are therefore excellent vaccine candidates for use in an approach to prevent severe disease. A key question, however, is whether their extensive diversity precludes them from forming components of the malaria vaccines of the future?