Improved retroviral suicide gene transfer in colon cancer cell lines after cell synchronization with methotrexate.
Cancer gene therapy by retroviral vectors is mainly limited by the level of transduction. Retroviral gene transfer requires target cell division. Cell synchronization, obtained by drugs inducing a reversible inhibition of DNA synthesis, could therefore be proposed to precondition target cells to retroviral gene transfer. We tested whether drug-mediated cell synchronization could enhance the transfer efficiency of a retroviral-mediated gene encoding herpes simplex virus thymidine kinase (HSV-tk) in two colon cancer cell lines, DHDK12 and HT29. Synchronization was induced by methotrexate (MTX), aracytin (ara-C) or aphidicolin. Gene transfer efficiency was assessed by the level of HSV-TK expression. Transduced cells were driven by ganciclovir (GCV) towards apoptosis that was assessed using annexin V labeling by quantitative flow cytometry. DHDK12 and HT29 cells were synchronized in S phase with MTX but not ara-C or aphidicolin. In synchronized DHDK12 and HT29 cells, the HSV-TK transduction rates were 2 and 1.5-fold higher than those obtained in control cells, respectively. Furthermore, the rate of apoptosis was increased two-fold in MTX-treated DHDK12 cells after treatment with GCV. Our findings indicate that MTX-mediated synchronization of target cells allowed a significant improvement of retroviral HSV-tk gene transfer, resulting in an increased cell apoptosis in response to GCV. Pharmacological control of cell cycle may thus be a useful strategy to optimize the efficiency of retroviral-mediated cancer gene therapy.


Effect of two different meals on bioavailability of nilvadipine in healthy volunteers.
The effect of two different meals on the bioavailability of nilvadipine, a new antihypertensive and antianginal drug, was examined in 16 healthy male volunteers in two separate studies. In each study of eight subjects in a Latin-square, two-way crossover design, two groups of four subjects each were given a single 6-mg oral dose of nilvadipine after overnight fasting or 30 minutes after a 464- or 748-kcal meal. There were no significant differences in the area under the plasma concentration-time curve or the maximum plasma concentration between the fasting and fed states for either meal. Although the time to reach the maximum plasma concentration was about the same after a 464-kcal meal and after fasting, it increased slightly but significantly after a 748-kcal meal, indicating possible delay in drug absorption after meals. These studies showed that the extent of bioavailability of nilvadipine appears to be little affected in the presence of food. Although a possible delay in the onset of absorption would occur, such a delay may not have any therapeutic importance in chronic therapy.


Placing joint hypermobility in context: traits, disorders and syndromes.
Joint hypermobility (JHM) is a common physical trait. It may occur alone or in combination with musculoskeletal (MSK) pain, outside or within more complex phenotypes. Hypermobility spectrum disorders (HSD) are diagnosed in individuals with JHM and related MSK pain, when an alternative diagnosis cannot be identified. Conversely, the Ehlers-Danlos syndrome (EDS) encompasses a group of rare hereditary connective tissue disorders featuring JHM along with other pleiotropic manifestations. The 2017 EDS Classification identifies 13 different subtypes. Hypermobile EDS (HEDS) is the only EDS variant still lacking a confirmatory test. Literature was reviewed searching for the most relevant papers related to key arguments. Particular attention was focused on papers published after the 2017 Classification. Definition, epidemiology, assessment tools and patterns of JHM are presented. The morbid nature of the 2017 EDS Classification and of the 'spectrum' is also illustrated. We discuss current limitations and disagreements concerning the 'spectrum', HSD and HEDS. In the clinical context, elucidation of the pathophysiology of pain related to JHM should develop in parallel with the analysis of pleiotropic manifestations of syndromes with JHM. Future challenges concerning classification, nosology, diagnosis and management of JHM, EDS and related disorders are discussed.


Antigenicity of polypeptides: immunological unresponsiveness to copolymers of gamma-amino acids.
Immunological tolerance toward three synthetic random copolymers of the alpha-amino acids, glutamic, lysine, alanine, and tyrosine, was produced by a single injection of the polymers into newborn rabbits. The tolerant state could be extended by an additional intravenous injection of antigen. Repeated injections of the polymer in adjuvant mixture could "break" the tolerant state.


Age does not impact risk for urethroplasty complications after tubularized incised plate repair of hypospadias in prepubertal boys.
Patients often present before or after the recommended age of 6-18 months for hypospadias repair. Reports indicate complications may increase when repair is delayed past 6-12 months of age. We questioned if age was an independent risk for urethroplasty complications (UC). A prospectively maintained database of consecutive patients undergoing tubularized incised plate (TIP) repair was queried for age at surgery, primary or reoperative TIP, meatal location, glansplasty suture, and learning curve. The presence of UC (fistula, dehiscence, stricture, meatal stenosis) was analyzed with logistic regression. TIP repairs were performed for 669 consecutive prepubertal patients aged 3-144 months (mean 17.1, SD 22.5). Original meatal location was distal in 540 (80.7%), midshaft in 50 (7.5%), and proximal in 79 (11.8%). Reoperative TIP occurred in 73 (10.9%). UC occurred in 77 (11.5%). Reoperative TIP (OR 3.07, 95% CI 1.54-6.13) and meatal location (OR 1.79, 95% CI 1.34-2.40) were the only independent risk factors for UC. Neither younger nor older age increased risk for UC. Our data from consecutive TIP repairs in prepubertal children indicate age at surgery does not increase odds of UC. Surgery can be performed any time after 3 months (in full-term, healthy boys) without raising the rate of UC.


Harnessing the CRISPR-Cas13d System for Protein Detection by Dual-Aptamer-Based Transcription Amplification.
CRISPR-based biosensing technology has been emerging as a revolutionary diagnostic tool for many disease-related biomarkers. In particular, RspCas13d, a newly identified RNA-guided Cas13d ribonuclease derived from Ruminococcus sp., has shown great promise for accurate and sensitive detection of RNA due to its RNA sequence-specific recognition and robust collateral trans-cleavage activity. However, its diagnostic utility is limited to detecting nucleic-acid-related biomarkers. To address this limitation, herein we present a proof-of-concept demonstration of a target-responsive CRISPR-Cas13d sensing system for protein biomarkers. This system was rationally designed by integrating a dual-aptamer-based transcription amplification strategy with CRISPR-Cas13d (DATAS-Cas13d), in which the protein binding initiates in-vitro RNA transcription followed by the activation of RspCas13d. Using a short fluorescent ssRNA as the signal reporter and cardiac troponin I (cTnI) as the model analyte, the DATAS-Cas13d system showed a wide linear range, low detection limit, and high specificity for the detection of cTnI in buffer and human serum. Thanks to the facile integration of various bioreceptors into the DATAS-Cas13d system, the method could be adapted to detecting a broad range of clinically relevant protein biomarkers, and thus broaden the medical applications of Cas13d-based diagnostics.


9, 11-Azo-12-oxa-15-hydroxyprostanoic acid: a potent thromboxane synthetase inhibitor and a PGH2/TxA2 receptor antagonist.
9,11-Azo-13-oxa-15-hydroxyprostanoic acid (AOHP) is shown to be a potent inhibitor of thromboxane synthetase with an IC50 at about 10(-6)M. It also blocked the agonist actions of 9,11-epoxymethano-PGH2 (IC50 9 x 10(-7) M) and TxA2 (IC50 2.4 x 10(-6) M) on human platelet-rich plasma indicating that it also is a PGH2/TxA2 receptor blocker.


Association of Vitamin D Deficiency and Vitamin D Receptor Gene Polymorphisms with Type 1 Diabetes Risk: A South Indian Familial Study.
Vitamin D is a potent immune modulator and is associated with autoimmune diseases, predominantly Type 1 diabetes (T1D). Vitamin D status and its receptor gene polymorphisms in T1D are not yet investigated in the South Indian population. The present study focuses on exploring the significance of vitamin D levels and Vitamin D receptor (VDR) gene polymorphisms with the risk of T1D in the South Indian population. A total of 120 T1D patients along with 214 unaffected first-degree relatives (FDRs) were included in this study. Genotyping of VDR polymorphisms at four different loci (FokI- F/f, BsmI- B/b, TaqI- T/t, and ApaI- A/a) was assessed through the ARMS-PCR method. Serum vitamin D levels were measured in 98 T1D patients and 75 age and sex-matched siblings by the ELISA method. Vitamin D deficiency (VDD) was observed in a higher proportion of T1D patients than in controls (52% vs. 32%; p<0.03). The frequency of the FokI-FF genotype was significantly higher (OR=1.66; p<0.03) in T1D patients conferring a susceptible association with the disease. Nevertheless, the increased frequency of heterozygous Ff genotype (OR=0.57; p<0.02) among controls may confer a protective association with T1D. Further, the TDT analysis revealed over-transmission of ApaI-A (T: U=15/5; p<0.006) and BsmI-B alleles (T: U=17/5; p<0.01) and under-transmission of BsmI-b/ApaI-a/TaqI-T haplotype (T: U=5.4/14.4; p=0.04) from parents to T1D patients. The present study concludes that VDD is a major contributing risk factor to T1D development in the South Indian population. Furthermore, the FokI-FF genotype, BsmI-B, and ApaI-A alleles are positively associated with T1D risk. Whereas, the FokI-Ff genotype and BsmI-b/ApaI-a/TaqI-T haplotype are negatively associated with the disease.


Low-density granulocytes are related to shorter pregnancy duration but not to interferon alpha protein blood levels in systemic lupus erythematosus.
An increased risk of pregnancy complications is seen in women with systemic lupus erythematosus (SLE), but the specific immunopathological drivers are still unclear. Hallmarks of SLE are granulocyte activation, type I interferon (IFN) overproduction, and autoantibodies. Here we examined whether low-density granulocytes (LDG) and granulocyte activation increase during pregnancy, and related the results to IFNα protein levels, autoantibody profile, and gestational age at birth. Repeated blood samples were collected during pregnancy in trimesters one, two, and three from 69 women with SLE and 27 healthy pregnant women (HC). Nineteen of the SLE women were also sampled late postpartum. LDG proportions and granulocyte activation (CD62L shedding) were measured by flow cytometry. Plasma IFNα protein concentrations were quantified by single molecule array (Simoa) immune assay. Clinical data were obtained from medical records. Women with SLE had higher LDG proportions and increased IFNα protein levels compared to HC throughout pregnancy, but neither LDG fractions nor IFNα levels differed during pregnancy compared to postpartum in SLE. Granulocyte activation status was higher in SLE relative to HC pregnancies, and it was increased during pregnancy compared to after pregnancy in SLE. Higher LDG proportions in SLE were associated with antiphospholipid positivity but not to IFNα protein levels. Finally, higher LDG proportions in trimester three correlated independently with lower gestational age at birth in SLE. Our results suggest that SLE pregnancy results in increased peripheral granulocyte priming, and that higher LDG proportions late in pregnancy are related to shorter pregnancy duration but not to IFNα blood levels in SLE.


Systemic to pulmonary fistulas in Hodgkin's disease.
Systemic to pulmonary fistula is an unusual entity and its association with hematological diseases like Hodgkin's is extremely rare. We present a case of a 26-year-old woman with a thoracic mass and large arteriovenous fistula. The diagnosis of Hodgkin's disease, nodular sclerosis, was obtained by a biopsy of the supraclavicular lymph node. We achieved an excellent response after combined treatment with chemotherapy, radiotherapy and arterial embolization with complete disappearance of the mass.


Differences in prevalence and severity of coronary artery disease by three metabolic syndrome definitions.
People with metabolic syndrome (MS) are at increased risk for cardiovascular disease-associated morbidity and all-cause mortality. However, predicting the severity of coronary artery disease (CAD) according to different MS definitions is controversial. This study explores the prevalence and severity of CAD according to different MS definitions, focusing on their predictive value. We enrolled 690 Taiwanese adults with CAD in this study. Coronary artery lesions were classified by the modified American Heart Association-American College of Cardiology grading system. MS was diagnosed in 3 groups of randomly assigned subjects according to World Health Organization (WHO), International Diabetes Federation (IDF), and revised Adult Treatment Panel III (rATP III) criteria. More MS components and more severe CAD were identified in the WHO group. Prevalence of MS in the WHO, IDF, and rATP III groups was 33.9%, 47.4%, and 52.8%, respectively (P < 0.001). Mean lesion and culprit vessel numbers were slightly higher in WHO-group patients. The WHO group had more complex stenoses with type C classification than did the IDF and rATP III groups (32.7%, 26.3%, and 28.1%; P = 0.041) and significantly more calcified stenoses (21.5%, 16.2%, and 16.4%; P = 0.027). In a comparison of CAD severity by complex morphology lesions, the area under the receiver operating characteristic curve was higher in the WHO group (0.552; 95% confidence interval, 0.509-0.595; P = 0.019). Compared with IDF and rATP III definitions, the WHO definition of MS has superior predictive value of CAD severity in Taiwanese patients.


Lung perfusion and aerosol distributions in preterm ventilated lambs.
The relative distributions of ventilation as measured by 99Tc-sulfur colloid aerosol deposition and pulmonary perfusion (measured with radiolabeled microspheres) were determined in 12 preterm lambs that were delivered at 138 days gestational age and ventilated for 4 hrs. To verify that unventilated lung segments in these lambs would have decreased perfusion, a balloon catheter was placed in a major bronchus either at birth or after 2 hrs of ventilation. This catheter prevented ventilation of 24.5 +/- 3.2% of the lung tissue. After 4 hrs of ventilation, the lambs were sacrificed and the lungs were divided into about 60 1-g pieces. Apart from the occluded, atelectatic segments, the lungs were visually well aerated with only 5.3 +/- 1.8% of the nonobstructed lungs being spontaneously atelectatic. There was a 66.5 +/- 0.07% decrease in blood flow to the area of lung made atelectatic by the balloon. The blood flow also was decreased to lung regions assessed to be spontaneously atelectatic. No 99Tc-sulfur colloid was recovered from balloon-occluded lung regions, and less 99Tc-sulfur colloid was found in the spontaneously atelectatic areas than in aerated lung regions. There were significant correlations (P less than 0.001) between pulmonary blood flow and aerosol recovery in each of the 12 animals. Premature lambs had a wide variability in ventilation and perfusion, but the relative ventilation to perfusion ratio was regulated to minimize the intrapulmonary shunt.


Orbital cellulitis: a rare complication after orbital blowout fracture.
To report the incidence of orbital cellulitis after orbital blowout fracture. Retrospective, noncomparative, interventional case series. All patients with orbital cellulitis and a history of recent orbital fracture. A medical record review of clinical history, imaging studies, and surgical and treatment outcome was performed. Resolution of orbital cellulitis and surgical and imaging findings. Four patients (3 male; mean age, 30 years [range, 4.5-58]) were treated for orbital cellulitis complicating orbital fracture. All patients had evidence of paranasal sinusitis before or after the orbital injury, and 2 also reported forceful nose blowing after sustaining orbital trauma. Although 3 patients received prophylactic oral antibiotics after the fracture, this failed to prevent infection. Sinusitis commenced 1 to 2 weeks before and as late as 5 weeks after orbital injury. All patients were treated with IV antibiotics. Two developed an orbital abscess that required surgical drainage; 1 patient improved after an endonasal maxillary antrostomy. One patient improved on IV antibiotics alone and underwent fracture repair at a later stage. These 4 patients represent 0.8% of all cases of orbital fractures treated in the study period. Orbital cellulitis is a rare complication of orbital fracture, and seems to be more common when paranasal sinus infection preexists or occurs within several weeks of the injury. Oral antibiotics given after the orbital injury may not prevent orbital cellulitis or abscess formation. Surgery may be required to drain orbital abscess or in nonresolving cellulitis to drain the paranasal sinuses. Fracture repair, if indicated, should be delayed, particularly if an alloplastic implant is used.


Thermodynamic estimation of minor element distribution between immiscible liquids in Fe-Cu-based metal phase generated in melting treatment of municipal solid wastes.
Waste electrical and electronic equipment (WEEE) has become an important target in managing material cycles from the viewpoint of not only waste management and control of environmental pollution but also resource conservation. This study investigated the distribution tendency of trace elements in municipal solid waste (MSW) or incinerator ash, including valuable non-ferrous metals (Ni, Co, Cr, Mn, Mo, Ti, V, W, Zr), precious group metals (PGMs) originated from WEEE (Ag, Au, Pd, Pt), and others (Al, B, Pb, Si), between Fe-rich and Cu-rich metal phases by means of simple thermodynamic calculations. Most of the typical alloying elements for steel (Co, Cr, Mo, Nb, Ni, Si, Ti, V, and W) and Rh were preferentially distributed into the Fe-rich phase. PGMs, such as Au, Ag, and Pd, were enriched in the Cu-rich phase, whereas Pt was almost equally distributed into both phases. Since the primary metallurgical processing of Cu is followed by an electrolysis for refining, and since PGMs in crude copper have been industrially recovered from the resulting anode slime, our results indicated that Ag, Au, and Pd could be effectively recovered from MSW if the Cu-rich phase could be selectively collected.


Giant right coronary artery aneurysm presenting with non-ST elevation myocardial infarction and severe mitral regurgitation: a case report.
Coronary artery aneurysms are seen in 1.5-5% of patients presenting for coronary angiography, but giant aneurysms, defined as being greater than 2 cm in diameter, are rare. Given the paucity of cases and limited experience in diagnosis and management of the disease, each case is a learning tool in itself. We report the rare case of a 78-year-old Caucasian man who presented to a peripheral emergency department with chest pain and was subsequently found to have a giant right coronary artery aneurysm. Following initial investigation and treatment he was referred to our hospital for definitive management. The case described illustrates one of the varied presentations and subsequent management of an ill-defined and heterogeneous disease process. Given the limited experience with giant aneurysms in the coronary circulation, this case provides valuable insight into the clinical presentation of the disease and gives an example of the management of the most recent such case at our hospital.


Complement Component C3 and Complement Factor B Promote Growth of Cutaneous Squamous Cell Carcinoma.
Cutaneous squamous cell carcinoma (cSCC) is one of the most common metastatic skin cancers with increasing incidence. We examined the roles of complement component C3 and complement factor B (CFB) in the growth of cSCC. Analysis of cSCC cell lines (n = 8) and normal human epidermal keratinocytes (n = 11) with real-time quantitative PCR and Western blotting revealed up-regulation of C3 and CFB expression in cSCC cells. Immunohistochemical staining revealed stronger tumor cell-specific labeling for C3 and CFB in invasive cSCCs (n = 71) and recessive dystrophic epidermolysis bullosa-associated cSCCs (n = 11) than in cSCC in situ (n = 69), actinic keratoses (n = 63), and normal skin (n = 5). Significant up-regulation of C3 and CFB mRNA expression was noted in chemically induced mouse cSCCs, compared to benign papillomas. Knockdown of C3 and CFB expression inhibited migration and proliferation of cSCC cells and resulted in potent inhibition of extracellular signal-regulated kinase 1/2 activation. Knockdown of C3 and CFB markedly inhibited growth of human cSCC xenograft tumors in vivo. These results provide evidence for the roles of C3 and CFB in the development of cSCC and identify them as biomarkers and potential therapeutic targets in this metastatic skin cancer.


Investigations on asymmetric transmittivity of optical devices and different diode-like behaviors.
This study theoretically proved that although reciprocal optical devices can show asymmetric transmittivity (AT) under controlled incident modes (i.e., conditional AT), they cannot guarantee AT with arbitrary incident light modes, whereas only nonreciprocal optical devices can possibly guarantee AT. Besides, the thermodynamics of both reciprocal and nonreciprocal optical devices were discussed to show that the second law of thermodynamics is valid anyway. Furthermore, the diode-like behaviors of optical and electronic devices were compared. Electrons are identical to electronic devices, so electronic devices could have asymmetric conductance regardless of electrons. In contrast, electromagnetic waves are different from optical devices as transmittivity of different modes can be different, so reciprocal optical devices showing conditional AT cannot guarantee AT when incident modes are arbitrary. The mathematical proof and characteristic comparisons between electronic and optical diodes, which are firstly presented here, should help clarifying the necessary nonreciprocity required for being optical diodes.


Identification and characterization of SarH1, a new global regulator of virulence gene expression in Staphylococcus aureus.
The global regulators agr (accessory gene regulator) and sarA (staphylococcal accessory regulator) have been reported to be both activators and repressors of virulence gene expression in Staphylococcus aureus. How the effector of the agr system, RNAIII, interacts with target gene promoters is unknown. SarA, on the other hand, is a DNA-binding protein, which binds to conserved DNA motifs immediately upstream of both positively and negatively regulated promoters. Here, we searched for additional regulators that could explain the differential effects of RNAIII and SarA. Four differently regulated genes (hla, alpha-toxin; hld, RNAIII; spa, protein A; ssp, serine protease) were analysed for binding of potential regulatory proteins to the corresponding promoter DNA fragments, linked to magnetic beads. One protein (29 kDa), with affinity for all four promoters, showed a high degree of similarity to SarA and was named SarH1 (Sar homologue 1). Expression of sarH1 was strongly repressed by sarA and agr. Analysis of hla, hld, ssp and spa mRNAs in sarH1, sarA and agr mutants, and in sarA/sarH1 and agr/sarH1 double mutants, revealed that sarH1 has a strong repressive effect on hla and an activating effect on spa transcription. SDS-PAGE analysis of secreted proteins from the different mutants showed that the production of several other exoproteins was affected by sarH1. In conclusion, we show that both the agr-dependent suppression of protein A production and the sarA-dependent stimulation of alpha-toxin production is mediated via a new regulator, SarH1, which belongs to a family of Sar homologues.


Open-field behavior in two models of genetic hypertension and the behavioral effects of salt excess.
In order to assess the relationship of behavior to blood pressure and salt intake, open-field behavior was studied in 123 rats of the spontaneously hypertensive (SHR), Wistar-Kyoto (WKY), Sprague-Dawley (SD), and Dahl resistant (DR) and sensitive (DS) strains. DS rats become hypertensive upon exposure to either high dietary salt or psychogenic stress, while DR rats remain normotensive. A circular photobeam open field was used under full room illumination. Three 5-min trials were given on consecutive days. Rats were tested with or without 5 days of an 8% NaCl diet. We found that DS rats were less active than DR rats, regardless of diet. In contrast, SHR rats were more active than WKY rats. However, DS and SHR rats were equally active. Thus, behavioral differences between these two models of hypertension are expressed by the normotensive control strains. The relative ranking of activity levels between strains was DR = SD greater than SHR = DS greater than WKY. High-salt-enhanced intertrial habituation, defined as the decrease in activity across trials (DS, 100%; SD, 82%; SHR, 90%; WKY, 1350%; but DR, -50%) as well as intratrial habituation, defined as the decrease from the first to the second half of the trial (14%, all subjects). Defecation was increased with high salt (DS, 975%; SD, 59%; SHR, 267%; WKY, 89%; but DR, 40%). These effects of high salt may reflect an increase in an emotionality factor. DR rats were largely resistant to the behavioral effects of salt. Total activity was positively correlated with blood pressure in hypertensive rats, r(42) = 0.33, p less than .01, but negatively correlated in normotensive rats, r(81) = -0.34, p less than .01. The proportion of total activity occurring in the first half of the trial for the initial test day was correlated with blood pressure in normotensive rats r(81) = 0.44, p less than .01. Dietary salt excess has behavioral as well as cardiovascular consequences.


Trichomonas vaginalis (TV) and human papillomavirus (HPV) infection and the incidence of cervical intraepithelial neoplasia (CIN) grade III.
The temporal relationship between cervical infection with Trichomonas vaginalis (TV) or human papillomavirus (HPV) and the incidence rate of cervical intraepithelial neoplasia grade three (CIN III) was examined in a cohort of 43,016 Norwegian women. From 1980 to 1989, a cervico-vaginal infection from TV and HPV was diagnosed cytologically in 988 and 678 women, respectively. During the 181,240 person-years of observation, 440 cases of CIN III/cervical cancer developed. The age-adjusted incidence rates (IR) of CIN III were 225 per 100,000 person-years among women with no cytologic evidence of infection, 459 among women with TV infection, and 729 among women with HPV infection. A multiple regression model yielded a relative rate (RR) of CIN III of 2.1 (95 percent confidence interval [CI] = 1.3-3.4) among women with TV infection and 3.5 (CI = 1.9-6.6) among women with HPV infection, compared with women with neither infection. As CIN can be misclassified as HPV infection, the entry Pap-smears of 10 women with HPV infection who later developed CIN III were re-examined. Excluding the four discordant cases with the corresponding person-years decreased the RR of CIN III to 2.1 (CI = 0.9-4.8). Our report demonstrates the limitations of studies that rely only on cytologic detection of HPV infection. Nevertheless, the results support the hypothesis that HPV is a causal factor for CIN III lesions, and also display an association between TV infection and cervical neoplasia.


Evaluation of recombinant glutathione transferase 26 kDa, thioredoxin-1, and endophilin B1 of Taenia solium in the diagnosis of human neurocysticercosis.
Neurocysticercosis caused by Taenia solium larvae is a neglected disease that persists in several countries, including Mexico, and causes a high disability-adjusted life year burden. Neuroimaging tools such as computed tomography and magnetic resonance imaging are the most efficient for its detection, but low availability and high costs in most endemic regions limit their use. Serological methods such as lentil lectin-purified glycoprotein enzyme-linked immunoelectrotransfer blot antibody detection and monoclonal antibody-based enzyme-linked immunosorbent assays for HP10 antigen detection have been useful in supporting the diagnosis of this disease. We evaluated three T. solium recombinant antigens: glutathione transferase of 26 kDa (Ts26GST); thioredoxin-1 (TsTrx-1), and endophilin B1 (TsMEndoB1) by EITB. These are antigenic proteins antigenic, abundant in excretion/secretion products of the parasite, and do not cross-react with homologous host proteins. Ts26GST and TsTrx-1 showed sensitivity of 79 and 88%, specificity of 86 and 97%, PPV of 83 and 97% and NPV of 82 and 91%, respectively, for neurocysticercosis diagnosis. The recombinant antigens allowed the diagnosis of 70% (Ts26GST) and 80% (TsTrx-1) of patients having only one cysticercus. Further studies on specific regions of these proteins could improve T. solium diagnostics.


Glucose production in response to glucagon is comparable in preterm AGA and SGA infants.
Low plasma glucose concentrations are more often detected in small-for-gestational-age (SGA) than in appropriate-for-gestational-age (AGA) infants. This is ascribed to impaired glucose production due to presumed lower liver glycogen stores in SGA infants. The change in glucose production induced by glucagon is considered to be an indicator of liver glycogen content. We compared the effect of glucagon on glucose kinetics in preterm AGA and SGA infants. In 5 AGA and 5 SGA preterm infants (postnatal age: 3-6 days) glucose production and gluconeogenesis were measured using stable isotopes immediately before and for 1 h after a bolus of glucagon. After glucagon the plasma glucose concentration and glucose production increased significantly over time (P<0.05 and P<0.0001, respectively). The changes were comparable between AGA and SGA infants. Glycogenolysis contributed 75-80% to the increase in glucose production. The increase in glucose production after glucagon was similar in preterm AGA and SGA infants, and mainly due to an increase in glycogenolysis. Based on the assumption that glycogenolysis is an indicator of liver glycogen content, our data do not support the hypothesis that liver glycogen content is lower in preterm SGA compared to AGA infants after the first postnatal day.


Catalpol protects primary cultured astrocytes from in vitro ischemia-induced damage.
Catalpol, an iridoid glycoside abundant in the roots of Rehmannia glutinosa, has been previously found to prevent the loss of CA1 hippocampal neurons and to reduce working errors in gerbils after ischemia-reperfusion injury. In the present study, we investigated the effects of catalpol on astrocytes in an ischemic model to further characterize its neuroprotective mechanisms. Primary cultured astrocytes exposed to oxygen-glucose deprivation (OGD) followed by reperfusion (adding back oxygen and glucose, OGD-R), were used as an in vitro ischemic model. Treatment of the astrocytes with catalpol during ischemia-reperfusion increased astrocyte survival significantly in a concentration-dependent manner, as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release and morphological observation. In addition, catalpol prevented the decrease in mitochondrial membrane potential, inhibited the formation of reactive oxygen species (ROS) and the production of nitric oxide (NO), decreased the level of lipid peroxide and the activity of inducible nitric oxide synthase (iNOS), and elevated the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and the content of glutathione (GSH). Our results suggest that catalpol exerts the most significant cytoprotective effect on astrocytes by suppressing the production of free radicals and elevating antioxidant capacity.


iTRAQ-based quantitative proteomic analysis of cultivated Pseudostellaria heterophylla and its wild-type.
Pseudostellaria heterophylla is an important traditional Chinese herbal medicine with vast clinical consumption because of its positive effects. To date, changes in the metabolite composition of P. heterophylla have been well documented; however, the molecular differences between cultivated P. heterophylla and its wild-type are still unknown. The aim of this study was to investigate differences in cultivated and wild P. heterophylla. Due to the lack of a genomic database, we used high-throughput transcriptomic and proteomic technologies to identify proteins in the herb. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS were used to detect statistically significant changes between cultivated and wild P. heterophylla. We detected 3775 proteins; 332 showed differential accumulations across two different ecotypes of P. heterophylla. 71 significant differential expressions of proteins were selected based on GO annotations, KEGG, STRING analysis, and expression level. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. The results indicated that the carbohydrate and cellular amino acid metabolisms in cultivated P. heterophylla were weaker than those in its wild-type; seven important proteins were found to regulate sucrose and amino acids. This will provide the basic information for exploring the cause of differences in secondary metabolites in different ecotypes of P. heterophylla and the protein mechanism of its quality formation. This study combined a transcriptome, proteome, and metabolism approach for analyzing differentially expressed proteins of cultivated and wild P. heterophylla and established the relationship between significantly differentially expressed proteins and differential chemical components in non-model plants. The results of proteomic analysis provide the basic information for exploring the quality forming process, which will demonstrate, and provide guidance for, the study of effective constituents of P. heterophylla and its quality formation mechanism.


Human disorganization complex, as a polytopic blastogenesis defect: a new case.
We describe a baby girl of 4,000 g and 55 cm with supernumerary, malformed, and partially duplicated lower limbs, malformed and partially duplicated pelvis, spina bifida, coccygeal dermal sinus, ectopic anus located in the right buttock, duplicated internal genitalia, rectovaginal fistula, ileal atresia, Meckel diverticulum, and various renal system anomalies. We think that this phenotype is a new case of disorganization in humans (DsH) and postulate that this condition constitutes a polytopic defect of the blastogenesis. In this case, the presence of a malformation pattern involving structures in different parts of the body and organs derived from all of the germ layers, suggests that the pathogenetic event most probably occurred during blastogenesis affecting various progenitors fields.


Characterisation of von Willebrand factor A1 domain mutants I1416N and I1416T: correlation of clinical phenotype with flow-based platelet adhesion.
Type 2M von Willebrand disease (VWD) results from mutations in the A1 domain of von Willebrand factor (VWF) that reduce its platelet-binding function. However, currently employed VWF functional static assays may not distinguish between clinical phenotype. Fifteen individuals from five kindreds with VWF-A1 domain mutations I1416T or I1416N, correlated with mild and moderate clinical phenotypes, respectively, were investigated. The mutations were reproduced by site-directed mutagenesis and expressed in HEK293T cells; functional studies of the recombinant mutants, including GPIbα binding using a flow-based assay, were performed. Plasma from all individuals demonstrated discordant reductions in VWF antigen and platelet-binding function in the presence of high-molecular-weight VWF multimers consistent with VWD type 2M. There was lowered expression and secretion of both mutants compared with wild type (WT) recombinant (r)VWF as well as a significant reduction in GPIbα binding. Binding to collagen was normal and electrophoretic analysis demonstrated a similar multimer distribution between the mutant proteins and wt-rVWF. GPIbα binding under flow was also significantly reduced for I1416N and I1416T rVWF. Impairment of GPIbα binding was more marked for I1416N rVWF than I1416T under both static and flow conditions: this was in spite of similar VWF:Ristocetin cofactor (RCo) activities in patient plasma and is consistent with a respective clinical phenotype. Our findings have established for the first time that I1416N and I1416T are responsible for a type 2M VWD phenotype and demonstrate that quantification of VWF function under shear stress may provide a more accurate measure of clinical severity than the static functional measurements in current diagnostic use.


Comparing the toxicity of iodinated X-ray contrast media on eukaryote- and prokaryote-based quantified microarray assays.
This study compared the toxicity mechanisms of four X-ray-based iodinated contrast media (ICM) on Escherichia coli (E. coli) and yeast microarray assays, aiming to determine the diverse toxicity mechanisms among different exposed organisms and the relationship between toxicity and their physical and chemical characteristics. The conventional phenotypic endpoint cytotoxicity and the change in reactive oxygen species (ROS) level were employed in conjunction with toxicogenomics to quantify changes in the gene/protein biomarker level in the regulation of different damage/repair pathways. The results showed that molecular toxicity endpoints, named transcriptional effect level index (TELI) and protein effect level index (PELI) for E. coli and yeast, respectively, correlated well with the phenotypic endpoints. Temporal altered gene/protein expression profiles revealed dynamic and complex damage/toxic mechanisms. In particular, compared with E. coli cells, yeast cells exposed to ICM exhibited significantly higher stress intensity and diverse stress types, resulting in stress or damage to the organism. The toxic mechanisms of ICM are concentration/property-dependent and relevant to the cellular structure and defense systems in prokaryotes and eukaryotes. In particular, the toxicity of ionic ICM is higher than that of non-ionic ICM, and eukaryotes are more susceptible than prokaryotes to ICM exposure.


Impact on the quality of remaining life of treatment with self-expandable metal prostheses in patients with inoperable esophago-cardial cancer. Results of a study of 107 consecutive patients treated with Ultraflex-type prostheses.
Self-expanding metal stents are a new alternative for palliation of esophagocardial malignancies. We evaluated the impact of these stents on the quality of remaining life in patients affected by inoperable esophago-cardial cancer. Between September 1992 and September 1997, 102 patients underwent implantation of self expanding metal stents for palliation of dysphagia due to esophagene or cardial cancer (76 patients), or for locally recurrent carcinoma after surgery (14 patients), lasertherapy (13 patients) or radiotherapy (5 patients). Stents were implanted under radiological and endoscopic control in patients under mild sedation. Successful stent implantation was achieved in 102/107 patients (95.3%). Early complications were observed in 4.9% and per-operative mortality was 1.96%. After stent implantation the dysphagia score improved from 3.0, on average, to 0.5, on average. Late complications were evidenced in 25.5%. Weight gain was evidenced in 24.5% and the performance status improved in 14.3%. The mean survival time was 6.9 months. Self expanding metal stents are an effective alternative for palliation of malignant dysphagia due to esophageal and cardial cancers.


SARS-CoV-2 infection of the placenta.
BACKGROUNDThe effects of the novel coronavirus disease 2019 (COVID-19) in pregnancy remain relatively unknown. We present a case of second trimester pregnancy with symptomatic COVID-19 complicated by severe preeclampsia and placental abruption.METHODSWe analyzed the placenta for the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through molecular and immunohistochemical assays and by and electron microscopy and measured the maternal antibody response in the blood to this infection.RESULTSSARS-CoV-2 localized predominantly to syncytiotrophoblast cells at the materno-fetal interface of the placenta. Histological examination of the placenta revealed a dense macrophage infiltrate, but no evidence for the vasculopathy typically associated with preeclampsia.CONCLUSIONThis case demonstrates SARS-CoV-2 invasion of the placenta, highlighting the potential for severe morbidity among pregnant women with COVID-19.FUNDINGBeatrice Kleinberg Neuwirth Fund and Fast Grant Emergent Ventures funding from the Mercatus Center at George Mason University. The funding bodies did not have roles in the design of the study or data collection, analysis, and interpretation and played no role in writing the manuscript.


Prolonged effects of cyclic AMP analogues of phosphatidylcholine biosynthesis in cultured rat hepatocytes.
The short- and long-term effects of cyclic AMP analogues and phosphodiesterase inhibitors on phosphatidylcholine biosynthesis in monolayer cultures of rat hepatocytes were investigated. All the compounds tested produced an inhibition of phosphatidylcholine biosynthesis for up to 6 h after addition to the hepatocyte medium. The reduced rate of lipid synthesis was a function of the concentration of cyclic AMP analogue and was independent of the concentration of choline in the medium. The proportion of choline oxidized to betaine was relatively unaffected. Choline was incorporated into hepatocytes by saturable and non-saturable mechanisms. Although the various cAMP analogues had different effects on choline uptake, chlorophenylthio-cAMP reduced uptake of choline by 28% for cells treated for 1.5-15 h. This analogue lowered the Vmax of the saturable component of choline transport by 3.6-fold. Prolonged incubation of the hepatocytes with cAMP analogues resulted in a reversal of the inhibition of phosphatidylcholine synthesis. After 15 h all the compounds tested stimulated the relative incorporation of [methyl-3H]choline into phosphatidylcholine. For hepatocytes incubated with chlorophenylthio-cAMP for 14-16 h, there was a 2.8-fold stimulation of the rate of phosphatidylcholine synthesis. The enzymes responsible for the conversion of choline into phosphatidylcholine were examined at various times after addition of the chlorophenylthio-cAMP to the hepatocyte medium. The reduced synthesis of phosphatidylcholine strongly correlated with inhibition of CTP:phosphocholine cytidylyltransferase activity. After 12 h of treatment with the analogue, the relative inhibition of the cytidylyltransferase activity was reversed.


Visual outcomes following vitrectomy and peeling of epiretinal membrane.
Visual outcomes of patients following vitrectomy and peeling of visually significant epiretinal membranes were assessed to determine the influence of specific perioperative factors and surgical complications on final visual acuity and functional vision. In an unmatched, consecutive surgical series, vitrectomy and membrane peeling were performed on 125 eyes of 123 patients with visually significant macular epiretinal membranes. Patients were followed for 6-36 months. Visual outcome measures included postoperative logMAR visual acuity, change in visual acuity and functional vision tasks evaluated by questionnaire. Perioperative factors including duration of symptoms, preoperative visual acuity, aetiology, membrane type and leakage on fundal fluorescein angiogram were correlated with final visual outcomes. Visual acuity improved by a mean of 0.31 A+/- 0.21 units (three lines of vision). In 104 cases (83%), visual acuity improved in patients by two lines or more, with 20 cases (16%) having unchanged acuity and one case (1%) having worse acuity. Ninety-three per cent of interviewed cases reported improvement in functional vision, especially reduction of distortion. Cataract was observed in 52 cases (52% of phakic eyes) postoperatively compared with 19 cases (19%) preoperatively. Postoperative visual acuity correlated with preoperative visual acuity. Patients with worse preoperative vision recorded greater visual improvement following surgery. No other perioperative factors were found to have a prognostic value in this study. Epiretinal membrane peeling improves vision in the majority of patients with significant symptoms, even if preoperative visual acuity is not substantially reduced. Surgery improves functional vision including metamorphopsia not measurable by visual acuity, and thus assessment of functional vision should be included in surgical case planning.


The role of ovarian sympathetic innervation in the control of estrous responsiveness in the rat.
This study examined the contribution of the superior ovarian nerve (SON) to estrous responsiveness and ovarian function in cycling rats. Section of the SON was carried out at 1100 on proestrus, and lordotic responsiveness was measured at 1500, 1700, and 2100 on that day and at 0900, 1200, and 1500 on the day of estrus. SON section decreased lordosis intensity significantly at 1500 and 1700 on proestrus and at 0900 on estrus. Pacing of sexual contacts with males was decreased at 2100 in nerve-sectioned (Nervx) animals when compared with sham-operated controls (SHAM). Serum progesterone (P) concentrations were significantly lower in Nervx animals than in Sham animals 30 min after surgery, but were not different between groups at 4.5 hr. Serum estradiol (E2) concentrations did not differ between groups at either time. In addition, Nervx and Sham groups did not differ on measures of pregnancy/pseudopregnancy initiation or on measures of ovarian function 10 days after surgery. These data suggest that the integrity of the SON is necessary for the display of full estrous responsiveness in cycling rats, and suggest that the acute decreases in serum P occurring as a consequence of SON section may be responsible for the deficits seen in Nervx animals.


Removal of emerging organic micropollutants via modified-reverse osmosis/nanofiltration membranes: A review.
Hazardous micropollutants (MPs) such as pharmaceutically active compounds (PhACs), pesticides and personal care products (PCPs) have emerged as a critical concern nowadays for acquiring clean and safe water resources. In the last few decades, innumerable water treatment methods involving biodegradation, adsorption and advanced oxidation process have been utilized for the removal of MPs. Of these methods, membrane technology has proven to be a promising technique for the removal of MPs due to its sustainability, high efficiency and cost-effectiveness. Herein, the aim of this article is to provide a comprehensive review regarding the MPs rejection mechanisms of reverse osmosis (RO) and nanofiltration (NF) membranes after incorporation of nanomaterials and also surface modification atop the PA layer. Size exclusion, adsorption and electrostatic charge interaction mechanisms play important roles in governing the MP removal rate. In addition, this review also discusses the state-of-the-art research on the surface modification of thin film composite (TFC) membrane and nanomaterials-incorporated thin film nanocomposite (TFN) membrane in enhancing MPs removal performance. It is hoped that this review can provide insights in modifying the physicochemical properties of NF and RO membranes to achieve better performance in water treatment process, particularly for the removal of emerging hazardous substances.


Modified contrast microinjection technique to facilitate chronic total occlusion recanalization.
To assess the efficacy and safety of the modified contrast microinjection technique to facilitate chronic total occlusion recanalization. The success rate of chronic total occlusion percutaneous coronary intervention (CTO-PCI) does not exceed 90% even in the most experienced centres. We have previously demonstrated that a large volume of contrast injected into the subintimal space can facilitate recanalization but is limited by a risk of serious complication. The aim of this study was to assess the application, efficacy and safety of a modified contrast microinjection technique that utilizes the injection of a much smaller volume of contrast in CTO-PCI. A retrospective analysis of patients in whom the modified microinjection technique was utilised in patients undergoing CTO-PCI at five tertiary centres was conducted. Of 1,192 patients who underwent CTO-PCI, the microinjection technique was used in 59 patients (4.7%). The majority of CTOs treated were in the right coronary artery (79.7%), were of high complexity and 35.6% of lesions had a least one previously failed PCI attempt. The modified microinjection technique was used for more than one indication in 7 (11.9%) of patients. The success rate was 81.4%. There were no procedural complications related to the use of the reported modified microinjection. Initial experience with the modified microinjection demonstrates that it can be performed safely, is reproducible, and is broadly applicable. Whilst not required for all CTO procedures it can be a helpful adjunctive tool to increase the likelihood of success especially in complex, or refractory cases, without compromising the safety of the procedure. © 2015 Wiley Periodicals, Inc.


A perivascular system releasing sirolimus prevented intimal hyperplasia in a rabbit model in a medium-term study.
The main complication of aortocoronary reconstruction with vein grafts is restenosis in the course of time. The aim was to assess the effect of a periadventitial polyester mesh releasing sirolimus on intimal hyperplasia of autologous grafts. We implanted v. jugularis ext. into a. carotis communis in rabbits. The vein graft was either intact, or was wrapped with a pure polyester mesh, or with a sirolimus-releasing mesh. Three and six weeks after surgery, the veins were subjected to standard histological staining and the thicknesses of the tunica intima, the media and the intima-media complex were measured. Wrapping the vein with a mesh releasing sirolimus or with a pure mesh decreased the thickness of the intima in comparison with a vein graft by 73 ± 11% or 73 ± 8% after 3 weeks, and by 73 ± 9% or 59 ± 12% after 6 weeks, respectively. Sirolimus-releasing meshes reduced the thickness of the media by 65 ± 9% and 20 ± 12% after 3 and 6 weeks. The thickness of the intima-media complex in grafts with sirolimus-releasing meshes decreased by 60 ± 6% and 30 ± 13% in comparison with pure PES meshes, after 3 and 6 weeks, respectively. A periadventitial polyester mesh releasing sirolimus has the potential to become an effective device in preventing vein graft restenosis.


Pharmacokinetic assessment of dacomitinib (pan-HER tyrosine kinase inhibitor) in patients with locally advanced head and neck squamous cell carcinoma (LA SCCHN) following administration through a gastrostomy feeding tube (GT).
Dacomitinib is an irreversible oral pan-HER tyrosine kinase inhibitor with antitumor activity demonstrated in patients with recurrent/metastatic (RM) SCCHN. A Phase I trial of dacomitinib with standard therapy in LA SCCHN is ongoing (NCT01737008). As enteral feeding is needed for many SCCHN patients, this study investigated the PK properties of dacomitinib when administered via GT (NCT01484847). Since patients with GT are difficult to recruit, this study also determined the feasibility of PK assessments using a unique design in LA SCCHN patients with GT, by giving a single dose of drug during their radiotherapy (co-administration with chemotherapy avoided). Eligible patients were given a single dose of crushed dacomitinib at 45 mg in water suspension via GT. All doses were administered in fasting state and supine position. PK samples were drawn prior to dose (t = 0), 30 min and 1, 2, 3, 4, 6, 12, 24, 48, 72, 96, 144, 168, 192 and 216 hrs post-dose, and analyzed by HPLC-MS/MS. PK parameters (mean [CV%]) of this study were compared with those of dacomitinib given orally using Student t test. Six patients with LA SCCHN patients were enrolled. The median age of patients was 54 years. Two different types of GT were used: 14 F Cope-loop tube (n = 3), 20 F PEG/disc retention tube (n = 3). PK study showed t1/2 of 58 h, Cmax of 17 ng/ml, Tmax of 8 h, AUC0-inf of 1185 ng*hr/ml, Vd/F of 3310 L and CL/F of 41 L/hr. Compared with oral dosing of intact immediate release (IR) tablets, GT administration resulted in 34 % reduction in Cmax and 33-44 % decrease in AUC (all p <0.05) (Jänne et al., Clin Cancer Res 2011). Such differences were not detected when compared with the PK properties of dacomitinib administered orally in aqueous suspension (Bello et al., Cancer Chemother Pharm 2013). These differences may be attributed to aqueous suspension of dacomitinib. Caution should be taken with GT administration of orally active small molecule targeted therapy. This study also demonstrated that PK trials in GT patients are feasible using novel designs.


Antibody against the amino terminus of alpha-actin inhibits actomyosin interactions in the presence of ATP.
Actomyosin interactions in the presence of ATP were examined by using site-specific antibodies directed against the first seven N-terminal residues on skeletal alpha-actin. Fab fragments of these antibodies (S alpha N Fab) inhibited effectively the actin-activated ATPase of myosin subfragment 1 (S-1) at both 5 and 25 degrees C. Binding experiments carried out in the presence of ATP at 5 degrees C revealed that the catalytic inhibition was related to the inhibition of S-1 binding to actin by Fab. At equimolar ratios of Fab to actin, the binding of S-1 to actin and the activated ATPase were inhibited by 75 and 82%, respectively. These results, when contrasted with the small effect of Fab on rigor actomyosin binding, suggest ATP-induced changes at the interface of actin and myosin.


Potency of carbapenems for the prevention of carbapenem-resistant mutants of Pseudomonas aeruginosa: the high potency of a new carbapenem doripenem.
The potencies of the carbapenems; doripenem (DRPM), meropenem (MEPM) and imipenem (IPM) in preventing the emergence of carbapenem-resistant mutants were examined in Pseudomonas aeruginosa strains. The carbapenems predominantly selected carbapenem-resistant mutants or carbapenem mutants with reduced susceptibilities that were specifically resistant to carbapenems and had arisen as a result of the reduced level of expression of the outer membrane protein with a molecular weight of about 48,000 (OprD). The potency of carbapenems in preventing the growth of the mutants differed for DRPM, MEPM and IPM. The isolation frequency of the mutant was examined on agar plates containing each of the carbapenems at a concentration of 1/2 or 1/4 MIC of each carbapenem for that mutant. Mutants were not selected on agar containing DRPM at a frequency of greater than 10(-9) per cell per generation, whereas mutants of each strain were selected on agar containing MEPM or IPM at frequencies of 10(-7) to 10(-9) per cell per generation. The drug concentrations and the drug concentration range for the selective increase of carbapenem resistant mutants in the broth culture containing each carbapenem differed for each carbapenem. DRPM exhibited both the lowest drug concentration and the narrowest range of drug concentration for selection of the carbapenem-resistant mutants. The results shown in this report indicated that DRPM exhibited the greatest ability to prevent the emergence of the mutant.


Long-Term Treatment with Losartan Attenuates Seizure Activity and Neuronal Damage Without Affecting Behavioral Changes in a Model of Co-morbid Hypertension and Epilepsy.
Over the last 10 years, accumulated experimental and clinical evidence has supported the idea that AT1 receptor subtype is involved in epilepsy. Recently, we have shown that the selective AT1 receptor antagonist losartan attenuates epileptogenesis and exerts neuroprotection in the CA1 area of the hippocampus in epileptic Wistar rats. This study aimed to verify the efficacy of long-term treatment with losartan (10 mg/kg) after kainate-induced status epilepticus (SE) on seizure activity, behavioral and biochemical changes, and neuronal damage in a model of co-morbid hypertension and epilepsy. Spontaneous seizures were video- and EEG-monitored in spontaneously hypertensive rats (SHRs) for a 16-week period after SE. The behavior was analyzed by open field, elevated plus maze, sugar preference test, and forced swim test. The levels of serotonin in the hippocampus and neuronal loss were estimated by HPLC and hematoxylin and eosin staining, respectively. The AT1 receptor antagonism delayed the onset of seizures and alleviated their frequency and duration during and after discontinuation of treatment. Losartan showed neuroprotection mostly in the CA3 area of the hippocampus and the septo-temporal hilus of the dentate gyrus in SHRs. However, the AT1 receptor antagonist did not exert a substantial influence on concomitant with epilepsy behavioral changes and decreased 5-HT levels in the hippocampus. Our results suggest that the antihypertensive therapy with an AT1 receptor blocker might be effective against seizure activity and neuronal damage in a co-morbid hypertension and epilepsy.


Improving Contractors' Participation of Resource Utilization in Construction and Demolition Waste through Government Incentives and Punishments.
This paper develops a simulation model for analyzing how government incentives and punishments improve contractors' participation in resource utilization of construction and demolition waste (RUCDW) based on system dynamics theory. The construction industry's long-term objective is to become more sustainable and resource-effective, and as part of this objective, generated construction and demolition waste should be recycled and resource utilized. However, most contractors have little willingness to engage in RUCDW because it increases their costs. The government thus plays a vital role in improving their participation in RUCDW through a range of educational tools such as advertisements, professional training, incentives, and punishments. Among these approaches, incentives and punishments are considered the most effective because they directly change project costs. We use the Vensim software package for numerical simulation and data collected from Suzhou, China are used to demonstrate and validate the developed model. Simulation results show that the government can improve contractors' participation in RUCDW through three kinds of incentives and punishments: (1) subsidizing RUCDW; (2) increasing landfill fees; and 3) issuing fines for illegal dumping. Comprehensive application of multiple policies has a stronger effect than single policies. The established model is therefore a valuable tool for assessing the dynamic effects of government incentives and punishments on RUCDW ahead of implementation, which can provide guidance for policymakers.


Dynamic correlations between hemodynamic, metabolic, and neuronal responses to acute whole-brain ischemia.
Cerebral ischemia sets off a cascade of neuronal and metabolic responses to preserve brain viability. An understanding of the temporal evolution of these changes during and after ischemia, and their correlation with hemodynamic changes, is essential. In this study, a 12-min whole-brain ischemia based on the four-blood-vessel occlusion model was employed in rats. Using a high-temporal-resolution simultaneous (1)H-(31)P MRS acquisition sequence at 9.4 T, we investigated dynamic occlusion and reperfusion responses in cerebral lactate (Lac), phosphocreatine (PCr), adenosine triphosphate (ATP), pH, and blood oxygenation level dependence (BOLD), together with changes in neuronal field potential activity. We reveal tightly coupled dynamics between hemodynamic, metabolic, and neuronal responses to ischemia. Neuronal activity, BOLD, PCr, Lac, and pH changed immediately following occlusion, indicating reduced energy substrates and consumption, and increased glycolysis to maintain cellular ATP levels, which started to decrease 2.2 min after the onset of occlusion. ATP stores were then gradually consumed to maintain a minimum housekeeping neuronal activity level. By correlating dynamic changes of brain activity, BOLD, and energy metabolism, new insights into the brain's survival ability and mechanisms during an acute ischemic attack from the perspectives of cerebral metabolism, neuroenergetics, and neuronal activity were gained.


Aural Hematoma Secondary to Hemangiosarcoma in a Domestic Cat-Case Report.
A 7-year-old, white, domestic short hair, female cat was presented with an aural hematoma of the right pinna, which had been partially resected a year previously. A 3 × 4 cm mass, macroscopically similar to an auricular hematoma, was visible on the convex surface and a smaller vascular lesion was present on the ear margin. Cytological examination of the smaller mass was suggestive of hemangiosarcoma, and a diagnosis confirmed by histopathologic and immunohistochemical tests. Complete pinnectomy with a wide margin was performed and during the 3-year follow-up, the cat had no local recurrence or metastasis. We conclude that hemangiosarcoma should be considered as a potential differential diagnosis in white-coated cats with an unusual clinical presentation of aural hematoma.


Effect of anaesthesia on growth hormone secretion in the domestic fowl (Gallus domesticus).
Chloral hydrate, Equithesin, Althesin, Rompun, Sagatal and ether anaesthesia lowered the concentration of plasma growth hormone in immature cockerels. Handling and serial bleeding of the conscious controls had no effect on the growth hormone concentration.


Evaluation of extraocular electrodes for a retinal prosthesis using evoked potentials in cat visual cortex.
To assess the efficacy of a device using extraocular electrodes as a retinal prosthesis by evaluating the responses evoked in the visual cortex to electrical stimulation. In anaesthetised cats, a lateral orbital dissection and ipsilateral parietal craniotomy was performed. Two extraocular retinal prosthesis (ERP) disc electrodes were sutured to the sclera on the lateral and superior aspects of the globe. Retinal stimulation was performed with charge-balanced constant-current pulses. Potentials evoked in the visual cortex were measured with a ball electrode placed on the lateral gyrus after removal of the dura. Stable attachment of the ERP electrodes to the globe was achieved with scleral sutures. Visual cortex responses were recorded with the electrodes in bipolar and monopolar configurations. The evoked response consisted of an early component with a peak around 8 ms, and a late component with a peak after 50 ms. Thresholds for evoking a response occurred at current intensities as low as 500 microA. Through extrapolation from evoked response amplitude data, thresholds as low as 300 microA were calculated. Cathodal monopolar stimulation demonstrated lower thresholds than anodal stimulation for evoking cortical responses. The ERP electrodes can be easily attached to the globe and are effective in electrically stimulating the retina, evoking responses in the primary visual cortex. Threshold charge-density was within safe limits for neural stimulation.


Xeroderma pigmentosum fibroblasts of the D group lack an apurinic DNA endonuclease species with a low apparent Km.
Apurinic DNA endonuclease activity from cultured human fibroblasts was resolved into two species by phosphocellulose chromatography. The species had sedimentation coefficients of 3.3 S and 2.8 S and apparent Km's for apurinic sites of 5 and 44 nM, respectively. The low Km species was absent from extracts of cell lines XP5BE, XP6BE and XP7BE of xeroderma pigmentosum complementation group D.


Tenofovir versus entecavir on the prognosis of hepatitis B virus-related hepatocellular carcinoma: a systematic review and meta-analysis.
Tenofovir (TDF) and entecavir (ETV) are first-line treatments for patients with chronic hepatitis B virus (HBV) infection. However, the effect of TDF versus ETV on the prognosis of HBV-related hepatocellular carcinoma (HCC) has not been fully clarified yet. PubMed, Embase and Web of science were searched up to March, 2021. Meta-analyses were performed for overall survival (OS), disease-free survival (DFS) and recurrence-free survival (RFS) to assess the effect of TDF versus ETV on the prognosis of HBV-related HCC. A total of 10 studies comprising 4706 Asian patients were included. The pooled results revealed that TDF was associated with better OS (adjusted HR = 0.50, 95% CI: 0.40-0.62; I2 = 36.0%, p = 0.167) and better RFS/DFS (adjusted HR = 0.70, 95% CI: 0.55-0.89, I2 = 71.9%, p = 0.002) than ETV in treatment of HBV-related HCC. Subgroup analysis revealed that OS benefit from TDF was generally consistent, except for patients who underwent non-surgical treatment for HCC. Subgroup analysis also indicated that TDF reduces the risk of late recurrence (HR = 0.41, 95% CI: 0.18-0.0.93; I2 = 63.0%, p = 0.067) rather than early recurrence (HR = 0.99, 95% CI: 0.64-1.52; I2 = 61.3%, p = 0.076). Compared with ETV, TDF has the advantage of improving OS and reducing late recurrence of patients with HBV-related HCC patients who underwent resection.


Perfect Hamming code with a hash table for faster genome mapping.
With the advent of next-generation sequencers, the growing demands to map short DNA sequences to a genome have promoted the development of fast algorithms and tools. The tools commonly used today are based on either a hash table or the suffix array/Burrow-Wheeler transform. These algorithms are the best suited to finding the genome position of exactly matching short reads. However, they have limited capacity to handle the mismatches. To find n-mismatches, they requires O(2n) times the computation time of exact matches. Therefore, acceleration techniques are required. We propose a hash-based method for genome mapping that reduces the number of hash references for finding mismatches without increasing the size of the hash table. The method regards DNA subsequences as words on Galois extension field GF(2²) and each word is encoded to a code word of a perfect Hamming code. The perfect Hamming code defines equivalence classes of DNA subsequences. Each equivalence class includes subsequence whose corresponding words on GF(2²) are encoded to a corresponding code word. The code word is used as a hash key to store these subsequences in a hash table. Specifically, it reduces by about 70% the number of hash keys necessary for searching the genome positions of all 2-mismatches of 21-base-long DNA subsequence. The paper shows perfect hamming code can reduce the number of hash references for hash-based genome mapping. As the computation time to calculate code words is far shorter than a hash reference, our method is effective to reduce the computation time to map short DNA sequences to genome. The amount of data that DNA sequencers generate continues to increase and more accurate genome mappings are required. Thus our method will be a key technology to develop faster genome mapping software.


Inhibition of CYP3A-mediated oxidation in human hepatic microsomes by the dietary derived complex phenol, gallic acid.
Plant polyphenols, such as gallic acid, have been reported to have a range of biological activities including antimutagenic effects. Previously, we reported that gallic acid (3,4,5-trihydroxybenzoic acid), an agent found in wine and tea, inhibits androstenedione 6beta-hydroxylase activity (Ki 70 microm), a cytochrome P450 (CYP3A) marker in human liver microsomes. The pre-incubation of gallic acid (100 microM) with human liver microsomes in the absence of NADPH, as compared with the presence of NADPH, before assay of androstenedione 6beta-hydroxylase activity significantly increased the inhibitory effects of the gallic acid (0.03 +/- 0.03 nmol (mg microsomal protein)(-1) min(-1) compared with 0.20 +/- 0.06 nmol (mg microsomal protein) (-1) min(-1) (P < 0.05)). The antioxidant ascorbic acid and the radical scavenger glutathione prevented this observed increase in inhibition. Removal of gallic acid-derived products from the incubation completely restored CYP3A activity. In contrast, the activities of CYP1A and CYP2E, and non-CYP mediated reductive microsomal 17beta-hydroxysteroid dehydrogenase activity were refractory to inhibition by gallic acid.


Sepsis causes neutrophil infiltration in muscle leading to muscle atrophy and weakness in mice.
Sepsis-induced muscle atrophy leads to prolonged physical dysfunction. Although the interaction of muscle atrophy and macrophage has been reported in sepsis, the role of neutrophils in muscle atrophy has not been thoroughly investigated. This study sought to investigate the long-term changes in muscle-localized neutrophils after sepsis induction and their possible role in sepsis. Sepsis was induced in seven-week-old male C57BL/6J mice 8-12 (cecal slurry [CS] model) via intraperitoneal injection of 1 mg/g cecal slurry. The percentage change in body weight and grip strength was evaluated. The tibialis anterior muscles were dissected for microscopic examination of the cross-sectional area of myofibers or Fluorescence-activated cell sorting (FACS) analysis of immune cells. These changes were evaluated in the following conditions: (1) Longitudinal change until day 61, (2) CS concentration-dependent change on day 14 at the low (0.3 mg/g), middle (1.0 mg/g), and high (2.0 mg/g) concentrations, and (3) CS mice on day 14 treated with an anti-Ly6G antibody that depletes neutrophils. Body weight and grip strength were significantly lower in the CS model until day 61 (body weight: 123.1% ± 1.8% vs. 130.3% ± 2.5%, p = 0.04; grip strength: 104.5% ± 3.8% vs. 119.3% ± 5.3%, p = 0.04). Likewise, cross-sectional muscle area gradually decreased until day 61 from the CS induction (895.6 [606.0-1304.9] μm2 vs. 718.8 [536.2-937.0] μm2, p < 0.01). The number of muscle-localized neutrophils increased from 2.3 ± 0.6 cell/mg on day 0 to 22.2 ± 13.0 cell/mg on day 14, and decreased thereafter. In terms of CS concentration-dependent change, cross-sectional area was smaller (484.4 ± 221.2 vs. 825.8 ± 436.2 μm2 [p < 0.001]) and grip strength was lower (71.4% ± 12.8% vs. 116.3% ± 7.4%, p = 0.01) in the CS High group compared with the control, with increased neutrophils (p = 0.03). Ly6G-depleted mice demonstrated significant increase of muscle cross-sectional area and grip strength compared with control mice (p < 0.01). Sepsis causes infiltration of neutrophils in muscles, leading to muscle atrophy and weakness. Depletion of neutrophils in muscle reverses sepsis-induced muscle atrophy and weakness. These results suggest that neutrophils may play a critical role in sepsis-induced muscle atrophy and weakness.


Multicenter evaluation of a new capillary blood prothrombin time monitoring system.
The analytical performance of the new capillary blood prothrombin time monitoring system CoaguChek was examined in a multicenter evaluation at six hospitals. The coefficients of variation of the INR obtained in the CoaguChek imprecision study were approximately 7% in the control plasma provided (within-run and day-to-day) and 4% in blood (within-run). The prothrombin times were ascertained in capillary blood (CoaguChek PT Test) and citrated venous plasma (Hepato Quick, Thromborel S) from 359 patients under oral anticoagulation therapy with phenprocoumon, acenocoumarin or warfarin. The agreement with the test results obtained with the comparison methods was acceptable versus Hepato Quick assay (n = 359; y = 1.23 x -0.49, r = 0.888) and good versus Thromborel S method (n = 359; y = 1.09 x -0.28, r = 0.895). A simplified assessment of all test results (n = 795) in a nine-field comparison table showed a concordance with the comparison methods of more than 80 (Hepato Quick: 81%, Thromborel S: 83%). The concordance between Hepato-Quick and Thromborel S was slightly higher (88%). Its good analytical performance and convenient handling recommend the CoaguChek system as a suitable system for decentralized prothrombin time testing.


Comparison between different D-Dimer cutoff values to assess the individual risk of recurrent venous thromboembolism: analysis of results obtained in the DULCIS study.
D-dimer assay, generally evaluated according to cutoff points calibrated for VTE exclusion, is used to estimate the individual risk of recurrence after a first idiopathic event of venous thromboembolism (VTE). Commercial D-dimer assays, evaluated according to predetermined cutoff levels for each assay, specific for age (lower in subjects <70 years) and gender (lower in males), were used in the recent DULCIS study. The present analysis compared the results obtained in the DULCIS with those that might have been had using the following different cutoff criteria: traditional cutoff for VTE exclusion, higher levels in subjects aged ≥60 years, or age multiplied by 10. In young subjects, the DULCIS low cutoff levels resulted in half the recurrent events that would have occurred using the other criteria. In elderly patients, the DULCIS results were similar to those calculated for the two age-adjusted criteria. The adoption of traditional VTE exclusion criteria would have led to positive results in the large majority of elderly subjects, without a significant reduction in the rate of recurrent event. The results confirm the usefulness of the cutoff levels used in DULCIS.


Immunoglobulin mutant library genetically screened for folding stability exploiting bacterial signal transduction.
A model repertoire of variants of immunoglobulin kappa variable domain REIv with different folding stabilities was generated by oligonucleotide-directed randomization of position 29, a key conserved residue of hypervariable loop 1. Fused to ToxR', the membrane-anchored cytoplasmic domain of the Vibrio cholerae ToxR transcription activator, different members of the library induce different levels of transcription from the ctx promoter in Escherichia coli. Differences in transcription activation correlate positively with folding stabilities of the corresponding REIv domains. Since conformationally stabilized REIv derivatives elicit a dark red colony phenotype on EMB-lactose indicator plates, this procedure constitutes a genetic screen for immunoglobulin folding stability.


Case of spontaneous haemothorax associated with an endodermal sinus tumour.
Spontaneous haemothorax is rare in infants. A case is reported of a nine month old infant who was found to have an endodermal sinus tumour.


Confirmation of high blood flow rates through 150 mu filter/high-flow tubing.
Many new products designed to assist in rapid blood infusion are appearing. Some highly touted and routinely used devices for intravenous (IV) infusion have recently been shown to be, at least in part, defective. A tubing with an in-line 150 mu filter (150 mu High-Flow Blood Filter; Saftifilter Blood Administration Sets; Cutter Biological, Berkeley, CA 94710) has recently been introduced to facilitate rapid blood transfusion. It is claimed that at least 8.5 units of blood can be rapidly run through each set before replacement is necessary. To test this under simulated clinical conditions, four sets of ten random units of outdated erythrocytes at 4 to 9 degrees C were each admixed with 250 mL 70 degrees C 0.9 NaCl and infused through the system under a constant 300 mmHg pressure. Two sets infused through unmodified tubing flowed at an average of 25 mL/sec (1500 mL/min) before there was an appreciable slowing of the flow rate. Two sets with 8 Fr catheters attached infused at an average of 22 mL/sec (1320 mL/min) before there was an appreciable slowing of the flow rate. Even after the flow slowed, the 9th and 10th units infused at an average greater than 10 mL/sec (600 mL/min). The tubing/filter exceeded the manufacturer's published claims. This tubing/filter appears to be one element that could be an effective component of a high-flow infusion system.


Chimerism occurs in thyroid, lung, skin and lymph nodes of women with sons.
Chimerism indicates the presence of cells from one individual in another. Pregnancy and blood transfusions are considered the main sources for chimerism. Chimeric cells have been attributed a pathogenic role in various autoimmune diseases. However, data on the occurrence of chimeric cells in normal organs are scarce. In order to gain insight into the possible pathogenic potential of chimeric cells in autoimmune disease, it is necessary to determine the prevalence of chimeric cells in organs not affected by autoimmune disease. In situ hybridization for the Y-chromosome was performed on organs obtained at autopsy of 51 women. We investigated 44 thyroid, 38 lung, 21 skin and 7 lymph node samples. All women had sons, and data from their blood transfusion histories were retrieved for at least 10 years before death. Slides were scored semi-quantitatively for chimerism as low (1-3 Y-chromosome-positive cells per slide), moderate (4-10 positive cells per slide) or high (more than 10 positive cells per slide). Y-chromosome-positive cells were found in 8 thyroid, 10 lung, 3 skin and 1 lymph node samples of 18 women. There was no association between the presence of chimeric cells and blood transfusion history. Most organs in which chimerism was present contained a small to moderate level. Thus, chimerism can occur in normal organs of women without autoimmune disease. Our results indicate that chimerism is not necessarily associated with disease.


Structural and functional evolution of 2',3'-cyclic nucleotide 3'-phosphodiesterase.
2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) is an abundant membrane-associated enzyme within the vertebrate myelin sheath. While the physiological function of CNPase still remains to be characterized in detail, it is known - in addition to its in vitro enzymatic activity - to interact with other proteins, small molecules, and membrane surfaces. From an evolutionary point of view, it can be deduced that CNPase is not restricted to myelin-forming cells or vertebrate tissues. Its evolution has involved gene fusion, addition of other small segments with distinct functions, such as membrane attachment, and possibly loss of function at the polynucleotide kinase-like domain. Currently, it is unclear whether the enzymatic function of the conserved phosphodiesterase domain in vertebrate myelin has a physiological role, or if CNPase could actually function - like many other classical myelin proteins - in a more structural role. This article is part of a Special Issue entitled SI: Myelin Evolution.


Continuous cultivation of the diatom Nitzschia laevis for eicosapentaenoic acid production: physiological study and process optimization.
The continuous cultures of the diatom Nitzschia laevis were performed at different dilution rates (D) and feed glucose concentrations (S(0)) to investigate cellular physiological responses and its production potential of eicosapentaenoic acid (EPA). Steady-state cell dry weight, residual glucose concentration, cell growth yield, specific glucose consumption rate, and fatty acid profiles were investigated within the range of D from 0.1 to 1.0 day(-1) (S(0) fixed at 20 g/L) and the range of S(0) from 5 to 35 g/L (D fixed at 0.3 day(-1)), respectively. The highest EPA productivity of 73 mg L(-1) day(-1) was obtained at D = 0.5 day(-1) and S(0) = 20 g/L. However, when the continuous culture achieved high productivities of EPA at certain dilution rates and feed glucose concentrations, glucose in the feed could not be consumed completely. Accordingly, the continuous culture was evaluated in terms of both EPA productivity (P) and glucose assimilation efficiency (E). The parameter eta, defined as the product of P and E, was used as an overall performance index. Since eta is a function of the two independent variables D and S(0), we employed a central composite design to optimize D and S(0) for the highest eta value. Based on the experimental results of the design, a second-order polynomial equation was established to represent the relationship between eta and D and S(0). The optimal values of D and S(0) were subsequently determined as 0.481 day(-1) and 15.56 g/L, respectively by the empirical model. The verification experiment confirmed the validity of the model. Under the optimal conditions, eta value reached 46.5 mg L(-1) day(-1), suggesting a considerably high efficiency of the continuous culture of N. laevis in terms of EPA production and glucose utilization.


Leveraging user's performance in reporting patient safety events by utilizing text prediction in narrative data entry.
Narrative data entry pervades computerized health information systems and serves as a key component in collecting patient-related information in electronic health records and patient safety event reporting systems. The quality and efficiency of clinical data entry are critical in arriving at an optimal diagnosis and treatment. The application of text prediction holds potential for enhancing human performance of data entry in reporting patient safety events. This study examined two functions of text prediction intended for increasing efficiency and data quality of text data entry reporting patient safety events. The study employed a two-group randomized design with 52 nurses. The nurses were randomly assigned into a treatment group or a control group with a task of reporting five patient fall cases in Chinese using a web-based test system, with or without the prediction functions. T-test, Chi-square and linear regression model were applied to evaluating the outcome differences in free-text data entry between the groups. While both groups of participants exhibited a good capacity for accomplishing the assigned task of reporting patient falls, the results from the treatment group showed an overall increase of 70.5% in text generation rate, an increase of 34.1% in reporting comprehensiveness score and a reduction of 14.5% in the non-adherence of the comment fields. The treatment group also showed an increasing text generation rate over time, whereas no such an effect was observed in the control group. As an attempt investigating the effectiveness of text prediction functions in reporting patient safety events, the study findings proved an effective strategy for assisting reporters in generating complementary free text when reporting a patient safety event. The application of the strategy may be effective in other clinical areas when free text entries are required.


Nonlinear PI controllers for continuous bioreactors using population balance models.
Continuous bioreactors are critical unit operations in many biological systems, but the unique modeling is very complicated due to the underlying biochemical reactions and the distributed properties of cell population. The scope of this paper considers a popular modeling method for microbial cell cultures by population balance equation models, and the control objective aims to attenuate undesired oscillations appeared in the nonlinear distributed parameter system. In view of pursuing the popular/practical control configuration and the lack of on-line sensors, an approximate technique by exploiting the "pseudo-steady-state" approach constructs a simple nonlinear control model. Through an off-line estimation mechanism for the system having self-oscillating behavior, two kinds of nonlinear PI configurations are developed. Closed-loop simulation results have confirmed that the regulatory and tracking performances of the control system proposed are good.


Teaching of primary health care in practice: a model using local health centres in undergraduate medical education.
In Finland primary health care has a long historical background. The local communities, the state and the church have at various times and places been responsible for primary health care during the last few centuries. In 1972, a major reform took place when a new Primary Health Care Act came into force. In the same year two new medical faculties, at Kuopio and Tampere Universities, began to educate undergraduate medical students. In both of these new medical schools special attention was focused on the teaching of primary health care. Today practical teaching, which takes place at a primary health care centre, forms an important part of medical education at Kuopio University. This teaching of undergraduate students is part of the regular duties of general practitioners and public health nurses in the primary health care centres of eastern Finland that have agreed to collaborate in the teaching programme. The main principles are presented for the teaching programme in primary health care at the University of Kuopio.


NMDA Receptor-Dependent LTD Requires Transient Synaptic Incorporation of Ca²⁺-Permeable AMPARs Mediated by AKAP150-Anchored PKA and Calcineurin.
Information processing in the brain requires multiple forms of synaptic plasticity that converge on regulation of NMDA and AMPA-type glutamate receptors (NMDAR, AMPAR), including long-term potentiation (LTP) and long-term depression (LTD) and homeostatic scaling. In some cases, LTP and homeostatic plasticity regulate synaptic AMPAR subunit composition to increase the contribution of Ca(2+)-permeable receptors (CP-AMPARs) containing GluA1 but lacking GluA2 subunits. Here, we show that PKA anchored to the scaffold protein AKAP150 regulates GluA1 phosphorylation and plays a novel role controlling CP-AMPAR synaptic incorporation during NMDAR-dependent LTD. Using knockin mice that are deficient in AKAP-anchoring of either PKA or the opposing phosphatase calcineurin, we found that CP-AMPARs are recruited to hippocampal synapses by anchored PKA during LTD induction but are then rapidly removed by anchored calcineurin. Importantly, blocking CP-AMPAR recruitment, removal, or activity interferes with LTD. Thus, CP-AMPAR synaptic recruitment is required to transiently augment NMDAR Ca(2+) signaling during LTD induction.


Descending connections between superior olivary and cochlear nuclear complexes in the cat studied by autoradiographic and horseradish peroxidase methods.
The descending pathways from the superior olivary complex (SO) to the cochlear nuclear complex (CN) were investigated in 58 cats using labelled aminoacid and horseradish peroxidase transport techniques. Descending connections were found coursing bilaterally through the trapezoid body (TB) and ipsilaterally in the intermediate and dorsal acoustic striae. The dorsolateral periolivary nucleus (DLPO) sends fibres through both the intermediate and dorsal striae, which are joined by others from the lateral preolivary nucleus (LPO). Both the latter nucleus and the medial preolivary nucleus (MPO) give rise to a bilateral descending projection which traverses TB. The distribution of these descending pathways within CN is described (although the technique did not permit precise synaptic identification). The possible implications of these pathways for response patterns at the level of CN are discussed.


An informatic workflow for the enhanced annotation of excretory/secretory proteins of Haemonchus contortus.
Major advances in genomic and associated technologies have demanded reliable bioinformatic tools and workflows for the annotation of genes and their products via comparative analyses using well-curated reference data sets, accessible in public repositories. However, the accurate in silico annotation of molecules (proteins) encoded in organisms (e.g., multicellular parasites) which are evolutionarily distant from those for which these extensive reference data sets are available, including invertebrate model organisms (e.g., Caenorhabditis elegans - free-living nematode, and Drosophila melanogaster - the vinegar fly) and vertebrate species (e.g., Homo sapiens and Mus musculus), remains a major challenge. Here, we constructed an informatic workflow for the enhanced annotation of biologically-important, excretory/secretory (ES) proteins ("secretome") encoded in the genome of a parasitic roundworm, called Haemonchus contortus (commonly known as the barber's pole worm). We critically evaluated the performance of five distinct methods, refined some of them, and then combined the use of all five methods to comprehensively annotate ES proteins, according to gene ontology, biological pathways and/or metabolic (enzymatic) processes. Then, using optimised parameter settings, we applied this workflow to comprehensively annotate 2591 of all 3353 proteins (77.3%) in the secretome of H. contortus. This result is a substantial improvement (10-25%) over previous annotations using individual, "off-the-shelf" algorithms and default settings, indicating the ready applicability of the present, refined workflow to gene/protein sequence data sets from a wide range of organisms in the Tree-of-Life.


Combining concurrent radiotherapy and immunotherapy for synergistic effects in recurrent endometrial cancer - A case report.
Endometrial cancer, also known as uterine cancer, is the second most common cancer affecting women globally and the fourth most prevalent in the United States (US). Treatment often involves a combination of surgery, radiotherapy and chemotherapy depending on the severity. In this case report, we present two patients with relapsed endometrial carcinomas, who responded positively to combined radiotherapy and immunotherapy followed by maintenance immunotherapy. Given the worsening prognoses associated with recurrent endometrial cancers, these two cases warrant the further exploration of the concurrent administration of immunotherapy and radiation therapy in the context of clinical trials.


Development of an equation for calculating vertebral shear failure tolerance without destructive mechanical testing using iterative linear regression.
Equations used to determine vertebral failure tolerances without the need for destructive testing are useful for scaling applied sub-maximal forces during in vitro repetitive loading studies. However, existing equations that use vertebral bone density and morphology for calculating compressive failure tolerance are unsuitable for calculating vertebral shear failure tolerance since the primary site of failure is the pars interarticularis and not the vertebral body. Therefore, this investigation developed new equations for non-destructively determining vertebral shear failure tolerance from morphological and/or bone density measures. Shear failure was induced in 40 porcine cervical vertebral joints (20 C3-C4 and 20 C5-C6) by applying a constant posterior displacement to the caudal vertebra at 0.15 mm/s. Prior to destructive testing, morphology and bone density of the posterior elements were made with digital calipers, X-rays, and peripheral quantitative computed tomography. Iterative linear regression identified mathematical relationships between shear failure tolerance, and morphological and bone density measurements. Along with vertebral level, pars interarticularis length and lamina height from the cranial vertebra, and inferior facet height from the caudal vertebra collectively explained 61.8% of shear failure tolerance variance. Accuracy for this relationship, estimated using the same group of specimens, was 211.9 N or 9.8% of the measured shear failure tolerance.


Could serum visfatin be a potential biomarker for postmenopausal breast cancer?
Previous studies have shown that visfatin is significantly elevated in patients with gastric carcinoma and postmenopausal breast cancer (PBC). We thus explored whether serum visfatin could be used as a potential diagnostic and prognostic tool for PBC, taking into account clinicopathological features, serum tumor markers, anthropometric and metabolic parameters. Serum visfatin, tumor marker CA 15-3, carcinoembryonic antigen, metabolic and anthropometric parameters were determined in 103 postmenopausal women with pathologically confirmed, incident invasive breast cancer, 103 controls matched on age and time of diagnosis, and 51 patients with benign breast lesions (BBL). Mean serum visfatin was significantly higher in cases than in controls and patients with BBL (p<0.001). In cases, visfatin was significantly associated with CA 15-3 (p=0.03), hormone-receptor status (p<0.001), lymph node invasion (p=0.06) but not with metabolic and anthropometric variables (p>0.05). Multivariable regression analysis revealed that absence of estrogen and progesterone receptors (ER-PR-) was the strongest significant determinant of serum visfatin (p<0.001) in cases adjusting for demographic, metabolic and clinicopathological features. Based upon receiver operator characteristic analysis, serum visfatin outperformed CA 15-3 only in discriminating between PBC cases with early cancer stage than those with late stage, and in differentiating particularly patients with ER-PR- breast tumors. Further prospective and longitudinal studies are needed to determine whether serum visfatin could be used as a prognostic tool in the armamentarium of PBC monitoring and management in conjunction with other biomarkers.


Evaluation of the Merz Hand Grading Scale After Calcium Hydroxylapatite Hand Treatment.
Measurement scales that quickly and rigorously evaluate the effectiveness of filler treatment in hands are important tools in clinical practice. The Merz Hand Grading Scale (MHGS) is used to grade the appearance of the dorsal hand. The MHGS has been validated for photographic and live assessment of the hands. To evaluate the sensitivity of the 5-point MHGS to detect clinically meaningful and aesthetically pleasing changes in hand appearance after treatment with a calcium hydroxylapatite (CaHA)-based dermal filler. The controlled 4-week study randomized 30 subjects (60 hands) 2:1 to a Treatment group (treatment at enrollment) or a Control group (treatment at end of study). Effectiveness was evaluated with live MHGS ratings and photographic assessments with the Global Aesthetic Improvement Scale (GAIS). At Week 4, all Treatment group subjects (20/20) achieved a ≥1-point improvement on the MHGS compared with 0/10 (0%) of the Control group (p < .0001). Subjects and treating physicians rated 92.5% (37/40) and 100% (40/40), respectively, of hands as at least "improved," using the GAIS. The MHGS is an appropriate and validated tool that clinicians can use to counsel patients and evaluate clinically meaningful and aesthetically pleasing changes after hand treatment with CaHA.


Radiographic detection and characteristic patterns of residual excess cement associated with cement-retained implant restorations: a clinical report.
Residual excess cement (REC) is a common complication of cement-retained prostheses and has been linked to periimplant disease. Removal of the cement residue may result in resolution of the issue if addressed early in the disease process. However, this is dependent upon the ability to locate and adequately remove the foreign material. This series of patient scenarios describes the ability to detect REC by using dental radiography. Characteristics related to cements and flow patterns specific to implants are addressed.


Glomerular filtration rate estimated by the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation in type 1 diabetes based on genomic ancestry.
Black individuals have a great risk of developing chronic kidney disease (CKD) that is associated with high morbimortality, so it is important to classify them into the correct renal function group. Some equations used to estimate glomerular filtration rate (eGFR) divide patients only into two categories: African Americans and non-African Americans. The CKD-EPI equation was tested only in African Americans, and not Black patients from other regions, and takes into consideration self-reported color-race instead of genomic ancestry (GA) to determine the use of the ethnic correction factor. So far, this equation has not been evaluated in admixed populations, such as the Brazilian, using the percentage of GA to decide to apply the correction factor. The purpose of our study was to compare, in patients with type 1 diabetes (T1D), the eGFR calculated without the use of the correction factor, with the values obtained using the correction factor in patients presenting 50% or more of African GA. This cross-sectional, multicenter study enrolled 1279 patients from all geographic regions of Brazil. The CKD-EPI equation was used and CKD was defined as eGFR < 60 ml/min. GA were inferred using a panel of 46 AIM-INDEL, afterwards patients presenting an African GA ≥ 50% were selected. Initially, all patients with African GA ≥ 50% (n = 85) were considered as non-African Americans when calculating the eGFR and afterwards the ethnic correction factor was applied to recalculate the eGFR. CKD was present in 23 patients and 56.5% of them were redefined as having normal renal function after using the correction factor, mainly women [11 of the 13 patients (84.6%)], with GFR between 52-59.3 ml/min. More than half of the patients in the study were reclassified to a normal renal function group, showing that GA may be an important tool to decide between the use of the ethnic correction factor in the CKD-EPI equation in a highly admixed population of patients with T1D. A large-scale study involving GA and eGFR in comparison to reference methods should be conducted to better establish whether or not the ethnic correction factor should be used in highly admixed populations.