CD8+ T cell subpopulations and pro-inflammatory cytokines in neuromyelitis optica spectrum disorder.
Our study aimed to investigate circulating CD8+ T cell subpopulations and pro-inflammatory cytokines in the neuromyelitis optica spectrum disorder (NMOSD). A total of 121 peripheral blood samples were obtained from 57 patients with NMOSD, 34 patients with multiple sclerosis (MS), and 30 sex- and age-matched healthy controls (HCs) for detection of CD8+ T cell subpopulations, including phenotypes of naïve (TN , CD62Lhi CD45RO- ), effector/memory (TE/M , CD62Llo CD45RO+ ), memory precursor (TMP , CD127hi KLRG1lo ), and short lived effector (TSLEC , CD127lo KLRG1hi ). In addition, 36 samples from 18 NMOSD, 12 MS, and 6 sex- and age-matched HCs for detecting pro-inflammatory cytokines (IFNγ and TNFα) using flow cytometry. Compared with HCs, we found significantly reduced CD8+ TN and increased CD8+ TE/M in both NMOSD and MS,while decreased CD8+ TMP was only observed in NMOSD. Patients treated with immunotherapy were associated with increased CD8+ TN and decreased CD8+ TE/M in NMOSD. Moreover NMOSD cohort showed significant higher proportions of IFNγ+ CD8+ T cells and proportions of TNFα+ CD8+ T cells than HC and MS cohorts. On the contrary, obviously decreased IFNγ and TNFα were found in NMOSD patients treated with immunotherapy. Furthermore, Multivariate linear regression analyses revealed that age was negatively correlated with CD8+ TN and TMP , and positively associated with TSLEC ; however, sex, EDSS scores and disease phase were not significantly associated with CD8+ T subpopulations. This current study provides an evidence that circulating CD8+ T cell with abnormal subpopulations and increased pro-inflammatory were associated with pathogenesis of autoimmune demyelinating disease of CNS, especially in NMOSD.


A simplified cuff technique for abdominal aortic transplantation in mice.
Allograft arteriopathy is still a leading cause of late organ failure. The aortic allograft model in mice has been used to study chronic rejection and has given useful information in the development of graft arteriosclerosis. However, the technical difficulties of small vessel anastomoses still continue to limit its widespread use. We introduce a new simple method for aortic transplantation in mice. The descending aorta or infrarenal aorta from the donor mouse was anastomosed to the infrarenal aorta using a cuff technique. Aortic transplantation was performed in 30 mice, 10 isografts and 20 allografts. No immunosuppression was administered, and the recipients were sacrificed at day 28. The grafts were histologically analyzed. Implantation of grafts could be completed in an average of 23 min. There was no technical failure in all 60 anastomoses. The overall survival rate was 93.3%. Histology of aortas revealed typical aspects of chronic rejection in the allografts at day 28. No significant lesion was observed in isografts. We have developed an innovative, stable, and simple aortic transplantation model in mice, which is useful for vascular research in transplantation and beyond.


Daily estimation of the severity of organ dysfunctions in critically ill children by using the PELOD-2 score.
Daily or serial evaluation of multiple organ dysfunction syndrome (MODS) scores may provide useful information. We aimed to validate the daily (d) PELOD-2 score using the set of seven days proposed with the previous version of the score. In all consecutive patients admitted to nine pediatric intensive care units (PICUs) we prospectively measured the dPELOD-2 score at day 1, 2, 5, 8, 12, 16, and 18. PICU mortality was used as the outcome dependent variable. The discriminant power of the dPELOD-2 scores was estimated using the area under the ROC curve and the calibration using the Hosmer-Lemeshow chi-square test. We used a logistic regression to investigate the relationship between the dPELOD-2 scores and outcome, and between the change in PELOD-2 score from day 1 and outcome. We included 3669 patients (median age 15.5 months, mortality rate 6.1%, median length of PICU stay 3 days). Median dPELOD-2 scores were significantly higher in nonsurvivors than in survivors (p < 0.0001). The dPELOD-2 score was available at least at day 2 in 2057 patients: among the 796 patients without MODS on day 1, 186 (23.3%) acquired the syndrome during their PICU stay (mortality 4.9% vs. 0.3% among the 610 who did not; p < 0.0001). Among the 1261 patients with MODS on day 1, the syndrome worsened in 157 (12.4%) and remained unchanged or improved in 1104 (87.6%) (mortality 22.9% vs. 6.6%; p < 0.0001). The AUC of the dPELOD-2 scores ranged from 0.75 (95% CI: 0.67-0.83) to 0.89 (95% CI: 0.86-0.91). The calibration was good with a chi-square test between 13.5 (p = 0.06) and 0.9 (p = 0.99). The PELOD-2 score on day 1 was a significant prognostic factor; the serial evaluation of the change in the dPELOD-2 score from day1, adjusted for baseline value, demonstrated a significant odds ratio of death for each of the 7 days. This study suggests that the progression of the severity of organ dysfunctions can be evaluated by measuring the dPELOD-2 score during a set of 7 days in PICU, providing useful information on outcome in critically ill children. Its external validation would be useful.


Using FirePlex™ Particle Technology for Multiplex MicroRNA Profiling Without RNA Purification.
Accuracy of miRNA profiling is enhanced when sample processing can be kept to a minimum, avoiding steps such as RNA purification that can introduce bias and inaccuracies. Here we describe a novel multiplex circulating miRNA assay that enables the profiling of up to 65 miRNAs of choice in the same well directly from plasma (including heparin plasma) or serum, with no need for RNA purification. The main component of the assay is FirePlex™ hydrogel particles, which enable the multiplex capture of miRNAs with picomolar sensitivity and high specificity. Results are obtained using conventional flow cytometry and easy to use software, which allows fast analysis and interpretation of the experimental data. This chapter provides methods to profile miRNAs with PCR sensitivity from as little as 10 μL of crude biofluid sample, or from less than 100 pg of purified RNA.


Importance of fibroblast growth factor receptor in neovascularization and tumor escape from antiangiogenic therapy.
Therapeutic inhibition of pathways involved in angiogenesis has become the standard of care in renal cell carcinoma (RCC). Most currently available antiangiogenic agents inhibit the vascular endothelial growth factor (VEGF) pathway. Although these drugs have produced exciting benefits, some tumors do not respond to these agents. In addition most if not all tumors that initially respond will eventually develop resistance. Tumor escape from antiangiogenic therapy may include various signaling pathways that are involved in angiogenesis, including the fibroblast growth factor (FGF) signaling pathway. Emerging preclinical data suggest that FGF and VEGF act distinctly and synergistically to promote tumor vascularization. The current review discusses the role of FGF signaling in resistance to anti-VEGF therapies and outlines potential therapeutic implications.


Prescription drug monitoring program design and function: A qualitative analysis.
Opioid-related overdose deaths are a major public health challenge. Forty-nine states have implemented Prescription Drug Monitoring Programs (PDMPs) that collect information about individuals' prescription medications. Little is known about state governments' implementation of PDMPs. We conducted semi-structured interviews with PDMP staff, law enforcement officials, and administrative agency employees to learn about their attitudes and experiences with PDMPs. From May 2015 to June 2016, we conducted 37 semi-structured interviews with state actors in four states. Questions focused on interviewees' perceptions about PDMP goals, home agency characteristics, and future PDMP initiatives. States were selected purposively. Interviewees were identified through purposive and snowball sampling. Interviewees identified key PDMP goals as: improve patient treatment decisions; influence prescribing practices; assist in the identification of "doctor shoppers"; and serve as a tool for law enforcement. Interviewees identified the following characteristics as key for a PDMP's home agency: regulatory and enforcement authority; intra- and inter-agency collaboration; and commitment to data quality and protection. Interviewees identified three promising areas for future PDMP efforts: data sharing and analysis; integration of PDMP data with electronic medical records; and training for current and potential PDMP users. Our findings reveal areas that states may want to prioritize, including improving prescribers' knowledge and use of the PDMP as well as fostering inter-agency collaborations that include PDMP staff. By capitalizing on these opportunities, state governments may improve the effectiveness of their PDMPs, potentially making them more useful tools to curb the morbidity and mortality associated with opioid use disorders.


The effect of organ culture on human trabecular meshwork.
The effect of organ culture on the trabecular meshwork was studied in a series of human eyes using a perfusion culture system. One eye of a pair was cultured while the fellow eye was immediately fixed in glutaraldehyde. Culture periods ranged from 2 to 28 days, during which a mean intraocular pressure of 27 mmHg +/- 15 was maintained. The perfused culture medium appeared to leave the eye via Schlemm's canal and collector channels, mimicking the in vivo situation. The trabecular meshwork was well maintained with this system in 21 of 25 eyes. Cells remained in position on trabecular lamellae, cellular organelles usually remained normal, and the lamellae remained intact. Culture-induced changes were noted, with some cells developing intracytoplasmic lipid vacuoles and other cells developing swollen mitochondria. In addition, scattered focal cell necrosis was observed, most often in the uveal meshwork. Meshwork cellularity (nuclei:solid tissue) was determined with an image analysis system; there was an average cell loss of 20-40% in the cultured eyes as compared to their respective fellow control eyes. Overall, organ culture appears to maintain the human trabecular meshwork for at least 28 days, allowing controlled experimental studies to be performed.


Yeast expression and characterization of SARS-CoV N protein.
The severe acute respiratory syndrome human coronavirus (SARS-CoV) nucleocapsid protein (N protein) is its most antigenic structural protein. The N protein gene has been cloned into a yeast expression vector pPIC9, transformed into Pichia pastoris strain GS115 and induced for expression by methanol. SDS-PAGE and Western blot showed that the N protein was expressed at a level of 3mg/ml of culture medium. Characterization by mass spectrometry, circular dichroism and fluorescence luminescence assays showed that the expressed N protein displayed a beta-sheet secondary structure in solution and it is stable in the pH range between 5.0 and 8.0. The P. pastoris-expressed N protein is believed to more closely resemble native SARS N protein than the bacterially expressed N protein.


Strain-rate-dependent material properties of human lung parenchymal tissue using inverse finite element approach.
Automobile crashes and blunt trauma often lead to life-threatening thoracic injuries, especially to the lung tissues. These injuries can be simulated using finite element-based human body models that need dynamic material properties of lung tissue. The strain-rate-dependent material parameters of human parenchymal tissues were determined in this study using uniaxial quasi-static (1 mm/s) and dynamic (1.6, 3, and 5 m/s) compression tests. A bilinear material model was used to capture the nonlinear behavior of the lung tissue, which was implemented using a user-defined material in LS-DYNA. Inverse mapping using genetic algorithm-based optimization of all experimental data with the corresponding FE models yielded a set of strain-rate-dependent material parameters. The bilinear material parameters are obtained for the strain rates of 0.1, 100, 300, and 500 s-1. The estimated elastic modulus increased from 43 to 153 kPa, while the toe strain reduced from 0.39 to 0.29 when the strain rate was increased from 0.1 to 500 s-1. The optimized bilinear material properties of parenchymal tissue exhibit a piecewise linear relationship with the strain rate.


Preparation of sulfatide mimicking oleic acid sulfated chitosan as a potential inhibitor for metastasis.
Sulfatide is associated with numerous health problems, affecting different parts of the human body, including the metastasis; however, the underlying mechanisms are yet to be fully elucidated. Sulfatide has been used to potential inhibitor for tumor cell metastasis. In the present study we synthesized oleic acid sulfated chitosan (OlcShCs). It shows structural similarity to sulfatide because of its functional groups (sulfate and fatty acyl chains). Chitosan has smart properties such as biocompatibility, biodegradability and non-toxicity. We have prepared oleic acid sulfated chitosan (OlcShCs) by chitosan modification to mimic sulfatide. Its structure was characterized by FT-IR, H-NMR, and thermogravimetric analysis. After characterization studies its antimicrobial, antifungal and cytotoxic properties were investigated. Oleic acid sulfated chitosan (OlcShCs) was tested for its anti-cancer potential against human cancer cell lines (HeLa (ATCC® CCL-2™)) for 24 h, 48 h and 72 h using the MTT assays. This new material which is soluble at physiological conditions, is a potential candidate for further metastasis inhibition investigations.


Cytomegalovirus reactivation and its clinical impact in patients with solid tumors.
Cytomegalovirus reactivation can be life threatening. However, little evidence on its incidence in solid cancers is available. Therefore our single center Cytomegalovirus polymerase chain reaction database with altogether 890 CMV positive blood serum samples of mainly hematological and oncological patients was retrospectively analyzed to examine the occurrence of Cytomegalovirus reactivation in patients with solid tumors, resulting in 107 patients tested positive for Cytomegalovirus reactivation. Seventeen patients with solid cancer and a positive CMV-PCR test were identified, of which eight patients had clinically relevant CMV disease and received prompt antiviral treatment. Five patients fully recovered, but despite prompt antiviral treatment three patients died. Among these three patients two had significant co-infections (in one case EBV and in the other case Aspergillus) indicating that that CMV reactivation was at least one factor contributing to sepsis. The patient with the EBV co-infection was treated in an adjuvant therapy setting for breast cancer and died due to Cytomegalovirus and Epstein-Barr virus associated pneumonia despite intensive therapy. The other two patients had progressive disease of an underlying pancreatic cancer at the time of CMV diagnosis. One patient died due to attendant uncontrollable Aspergillus pneumonia, the other patient most likely died independent from CMV disease because of massively progressive underlying disease. Cytomegalovirus reactivation and disease might be underestimated in routine clinical practice. In our retrospective analysis we show that approximately 50 % of our patients suffering from solid cancers with a positive Cytomegalovirus polymerase chain reaction also had clinically relevant Cytomegalovirus disease requiring antiviral therapy.


The effect of fluoroquinolones and antioxidans on biofilm formation by Proteus mirabilis strains.
Fluoroquinolones are a group of antibiotics used in urinary tract infections. Unfortunately, resistance to this group of drugs is currently growing. The combined action of fluoroquinolones and other antibacterial and anti-biofilm substances may extend the use of this therapeutic option by clinicians. The aim of the study was to determine the effect of selected fluoroquinolones and therapeutic concentrations of ascorbic acid and rutoside on biofilm formation by Proteus mirabilis. The study included 15 strains of P. mirabilis isolated from urinary tract infections in patients of the University Hospital No. 1 dr A. Jurasz in Bydgoszcz (Poland). The metabolic activity of the biofilm treated with 0.4 mg/ml ascorbic acid, 0.02 µg/ml rutoside and chemotherapeutic agents (ciprofloxacin, norfloxacin) in the concentration range of 0.125-4.0 MIC (minimum inhibitory concentration) was assessed spectrophotometrically. Both ciprofloxacin and norfloxacin inhibited biofilm formation by the tested strains. The biofilm reduction rate was correlated with the increasing concentration of antibiotic used. No synergism in fluoroquinolones with ascorbic acid, rutoside or both was found. The ascorbic acid and rutoside combination, however, significantly decreased biofilm production. Our research proves a beneficial impact of ascorbic acid with rutoside supplementation on biofilm of P. mirabilis strains causing urinary tract infections.


Prenatal diagnosis of a rare variant of hypospadias and review of the literature.
We present a case of a variant of hypospadias, diagnosed prenatally at 21 weeks' gestation, in which the penis and scrotum appeared normal but there was a cyst-like lesion and a urethrocutaneous fistula on the ventral side of the penis. Detailed sonographic examination and karyotyping confirmed this was an isolated lesion and helped the parents to decide on continuation of the pregnancy.


Is platelet-rich plasma right for your patient?
Many pro athletes claim that platelet-rich plasma has improved their performance and saved their careers. But is it right for your patients? And if so, which ones?


Phase I evaluation of a 40-kDa branched-chain long-acting pegylated IFN-alpha-2a with and without cytarabine in patients with chronic myelogenous leukemia.
Pegasys (PEG-IFN) is a modified form of recombinant human IFN-alpha-2a in which IFN-alpha is attached to a branched methoxypolyethylene glycol (PEG) moiety of large molecular weight (40 kDa). Such molecular modification results in sustained absorption after s.c. drug administration and a prolonged half-life. A phase I study of PEG-IFN was conducted in patients with chronic myelogenous leukemia (CML) who were previously treated with IFN-alpha to evaluate the effect of sustained exposure to IFN on patients with CML. Twenty-seven patients with long-term or IFN-refractory CML were enrolled in cohorts of three or six patients. PEG-IFN was given once weekly by s.c. injections starting at a dose of 270 microg/wk to a maximum dose of 630 microg/wk. Sixteen additional patients were treated with escalating doses of PEG-IFN ranging from 450 to 540 microg/wk in combination with two different schedules of low-dose cytarabine (1-beta-D-arabinofuranosylcytosine, ara-C). Serial venous blood samples were collected to evaluate the pharmacokinetic and pharmacodynamic characteristics of PEG-IFN in these patients. The dose-limiting toxicity (DLT) as defined by the protocol was not achieved at the highest dose tested of 630 mug/wk. With the addition of ara-C, the DLT was reached at 540 microg/wk. The safety profile was similar to that of unmodified IFNs. Of 27 patients treated with PEG-IFN, 14 (52%) achieved or maintained a complete hematologic response and three (11%) achieved a complete cytogenetic response. Among 16 patients treated with the combination of PEG-IFN and ara-C, 11 (69%) achieved or maintained complete hematologic remission and two (13%) achieved complete cytogenetic remission. The mean serum peak concentration (C(max)) of PEG-IFN increased from 9.4 to 28 ng/mL as the dose increased from 270 to 450 microg/wk, with no further increases in C(max) at higher dose levels. Serum concentration reached peak value starting about 48 hours after drug administration and was maintained at close to peak value throughout the dosing interval. The mean +/- SD area under the serum concentration-time curve (AUC) calculated after the first dose also increased from 1,022 +/- 694 to 3,343 +/- 2,728 ng hour/mL as dose was increased from 270 to 450 microg/wk, showing a dose-related increase in systemic exposure of PEG-IFN. As with C(max), the AUC did not increase at higher dose levels. The maximum induction (E(max)) of neopterin, the surrogate marker of the pharmacodynamic activity of PEG-IFN, increased from 120% to 361% over baseline values as the dose was increased from 270 to 540 microg/wk. On the once-weekly multiple dosing schedule, both the PEG-IFN and neopterin concentration seemed to reach steady state by week 5 and the steady-state values were maintained with chronic dosing over 6 months. Pegasys provided a significant advantage over standard IFN-alpha by enabling once-weekly dosing while maintaining acceptable safety, tolerability, and activity profiles. This branched 40-kDa PEG-IFN was well tolerated both as a monotherapy as well as in combination with ara-C. Demonstration of its sustained exposure, pharmacodynamic activity, hematologic response, and evidence of cytogenetic response in several patients in this limited study with either IFN-refractory or INF-intolerant patients provides a promise for further investigation in combination with new agents like imatinib.


Herpes simplex virus-1 replication in histiotypic rotation-mediated reaggregated murine brain.
Inbred mouse strains exhibit varying susceptibilities to severe herpes simplex virus (HSV)-1-related neurologic disease. HSV-1 replication was examined in neural tissue obtained from mouse strains susceptible (A/J, SJL), moderately resistant (Balb/c), or resistant (C57BL/6) to severe HSV-1 disease. Reaggregated brain cultures were prepared from mechanically dissociated fetal mouse brains maintained with constant rotation. The resulting aggregates each contain neurons, astrocytes, oligodendrocytes, and microglia. These were inoculated with 10(-2)-10(4) plaque-forming units (pfu) HSV-1 MacIntyre/aggregate. Aggregates and media were harvested at 24, 48, 72, and 96 hr post-inoculation (p.i.) and assayed for virus production by plaque titration. Brain cultures prepared from A/J, SJL, Balb/c, and C57BL/6 mice supported HSV-1 replication equally well: by 96 hr p.i., titers of 10(6) pfu/ml were produced by each strain at each inoculum. ID50s were similar for A/J and C57BL/6 cultures. There was no increased capacity for HSV-1 replication or for permissiveness for HSV-1 infection in histiotypic brain cultures from mouse strains susceptible to severe HSV-1 disease.


[Endoscope-assisted supraorbital keyhole approach for excision of suprasellar region tumor].
To study endoscope-assisted microsurgical technique with supraorbital keyhole approach for the treatment of suprasellar region tumor so as to maximize tumor removal and minimize operative-trauma. According to high resolution CT and MR images before operation, individual operation schemes were worked out. Skin incision was made in the eyebrow, and the diameter of supraorbital craniotomy was approximately 2 cm. Endoscope-assisted microsurgical technique was used to resect lesions in 16 consecutive patients. Total Tumors were removed in all of the 16 patients via the supraorbital keyhole approach. No postoperative complications occurred. Supraorbital keyhole approach may diminish tissue injury considerably and has proven to provide sufficient operating space in the suprasellar region for tumor removal.


Effects of acute normovolemic hemodilution on allogeneic blood transfusion & coagulation in orthognathic surgery: A randomized study.
Acute normovolemic hemodilution (ANH) is one of the important techniques predominantly used in cardiac, hepatic, and vascular surgery for decreasing allogeneic blood transfusion. However, the effect of ANH in orthognathic surgery has been rarely studied. Therefore, this study aims to assess the ANH-mediated reduction in the allogeneic red blood cell transfusion for orthognathic surgery patients. In this single-center study, 18-80 years old patients were recruited. Patients with hemoglobin ≥11 g/dL and normal coagulation function were randomly divided into ANH or standard treatment group. Ninety six patients underwent ANH, and 101 patients received standard treatment. No differences in demographic or major pre-operative characteristics were observed between the two groups. One patient in the ANH and three patients in the standard treatment group received allogeneic blood [3(2.97%) vs. 1(1.16%), control vs. ANH, p = .395]. Multivariate logistic regression analysis revealed that ANH treatment was not associated with transfusion of allogeneic blood (p = .763). After retransfusing autologous blood, PT and APTT in the ANH group significantly increased compared to standard treatment group (PT: -1.73 ± 1.09 vs. -2.15 ± 1.06, p = .035; APTT: -6.39 ± 5.76 vs. -8.16 ± 5.70, p = .031; control vs. ANH). No significant differences between the two groups were observed for changes in coagulation parameters at first postoperative day. However, platelet counts in the ANH group decreased compared to the standard group. No significant difference in major adverse outcomes was observed between the two groups. ANH did not reduce the incidence of allogeneic transfusion in patients undergoing orthognathic surgery.


In patients hospitalized for COVID-19, tocilizumab reduces mortality at 28 d.
WHO Rapid Evidence Appraisal for COVID-19 Therapies (REACT) Working Group; Shankar-Hari M, Vale CL, Godolphin PJ, et al. Association between administration of IL-6 antagonists and mortality among patients hospitalized for COVID-19: a meta-analysis. JAMA. 2021;326:499-518. 34228774.


Rapid micromeasurement of neutrophil exocytosis.
A simple method is described for the visual detection of neutrophil granule exocytosis. The microassay measures release of myeloperoxidase from azurophilic granules following neutrophil stimulation and can be accomplished in less than 30 min. The assay is sensitive, reproducible from day to day, and does not require separation of leukocytes from the reaction before enzyme assay. The procedure is also applicable to the rapid measurement of the suppression of degranulation by antiinflammatory agents.


Lutembacher's syndrome with small atrial septal defect diagnosed by transthoracic and transesophageal echocardiography that underwent mitral valve replacement.
Lutembacher's syndrome is a rare clinical combination of congenital ostium secundum atrial septal defect associated with acquired mitral valve stenosis (usually rheumatic). This unusual cardiac entity is difficult to diagnose clinically because each lesion alters the hemodynamics and clinical characteristics of the other. The resulting clinical manifestations depend chiefly on the size of the defect, the severity of the mitral stenosis, and the compliance of the right ventricle. We present a classic case of Lutembacher's syndrome and illustrate the pitfalls and advantages of echocardiography in the correct diagnosis of the syndrome.


Rapid flocculation and settling of positively buoyant microplastic and fine-grained sediment in natural seawater.
Interactions between microplastic (MP) and fine-grained suspended sediment in natural waters are important for the environmental fate of plastic particles. Estuaries are transitional areas between freshwater and open marine systems and are recognized as important accumulation zones for MPs. However, there is a knowledge gap on the processes driving the sedimentation of MPs in estuaries, especially with regard to positively buoyant MPs. Here we show from settling tube experiments that positively buoyant and non-spherical MP HDPE particles in different size-fractions (63-500 μm) and concentrations (1 and 5 mg l-1) rapidly flocculate and settle with natural fine-grained sediment in natural seawater. Our results demonstrate that flocculation is a key process for the vertical transport of MP in estuaries. The implication is that land-based sources of positively buoyant HDPE MP transported by rivers will likely settle and accumulate in estuarine environments and thereby increase the concentration of MP in the benthic zone.


Effects of serotonin agonists on lordosis, myoclonus, and cytoplasmic progestin receptors in guinea pigs.
Peripheral treatment with the serotonin releaser fenfluramine or the serotonin agonist quipazine abolished lordosis behavior in ovariectomized estradiol and progesterone-primed female guinea pigs. Quipazine was also effective when administered into a lateral cerebral ventricle. The lowest dose of fenfluramine that induced myoclonus (10 mg/kg) was higher than the dose needed to inhibit lordosis (5 mg/kg). Therefore, it appears that myoclonus and lordosis are differentially sensitive to serotonin agonists. The effects of quipazine on lordosis were time dependent. Quipazine had no effect on lordosis when given prior to the onset of sexual receptivity. These data suggest that serotonin agonists might be effective only when progesterone has had sufficient time to induce sexual receptivity. Quipazine did not affect cytoplasmic progestin receptors in brain areas involved in steroid hormone effects on lordosis. This finding, and the finding that quipazine had no effect on lordosis when given prior to the onset of sexual receptivity, suggest increased serotonin transmission does not interfere with estrogen priming or sensitivity of hypothalamic cells to progesterone.


Chemoprevention of colon carcinogenesis by concurrent administration of piroxicam, a nonsteroidal antiinflammatory drug with D,L-alpha-difluoromethylornithine, an ornithine decarboxylase inhibitor, in diet.
The effect of three levels of piroxicam and three levels of D,L-alpha-difluoromethylornithine (DFMO) fed individually and in combination during the postinitiation phase of carcinogenesis was studied in male F344 rats to generate a data base on the efficacy and synergistic and additive effects of these compounds as inhibitors of colon carcinogenesis. The maximum tolerated dose of DFMO was determined in male F344 rats and found to be 5000 ppm in the AIN-76A diet. Piroxicam at levels of 25, 75, and 150 ppm and DFMO at concentrations of 400, 1000, and 4000 ppm (20, 50, and 80% maximum tolerated dose) in AIN-76 diet were tested individually and in combinations. At 7 weeks of age, while the rats were consuming the control diet (AIN-76A), all animals except the vehicle (saline)-treated controls were given a single s.c. injection of azoxymethane (CAS: 25843-45-2) at a dose level of 29.6 mg/kg body weight to induce intestinal tumors. One week after azoxymethane injection, animals were transferred to their respective experimental diets containing piroxicam and DFMO. Fifty-six weeks after azoxymethane injection, all animals were necropsied and colon and small intestinal tumor incidences and multiplicity were compared among the various dietary groups. Feeding of diets containing 75 and 150 ppm piroxicam or 1000 and 4000 ppm DFMO significantly inhibited the incidence (percentage of animals with tumors) of colon adenocarcinomas compared to that of control diet. The multiplicity (number of tumors/rat) of adenocarcinomas was significantly inhibited in animals fed the 25, 75, and 150 ppm piroxicam or 400, 1000, and 4000 ppm DFMO diets. Results analyzed by the linear regression method suggested a dose-dependent inhibition in colon adenocarcinoma incidence with increasing levels of piroxicam or DFMO. The incidence and multiplicity of colon adenocarcinomas were significantly inhibited in animals fed the diets containing combinations of 25, 75, and 150 ppm piroxicam and 400, 1000, and 4000 ppm DFMO. Piroxicam and DFMO administered together had a stronger inhibitory effect than did those given individually. Piroxicam and DFMO when administered individually had no significant inhibitory effect on colon adenoma incidence and multiplicity; in contrast, combinations of these compounds significantly inhibited colon adenomas. No consistent differences were found in the incidence and multiplicity of small intestinal tumors among the dietary groups.(ABSTRACT TRUNCATED AT 400 WORDS)


Totally endoscopic aortic valve replacement with concomitant trans-aortic mitral valve repair for mitral regurgitation.
Minimally invasive aortic valve procedures through a hemi-sternotomy or a right anterior mini-thoracotomy have gained popularity over the last several years. Totally endoscopic aortic valve replacement (TEAVR) is an innovative and a less invasive (incision-wise) surgical aortic valve replacement technique. The operative steps of TEAVR have been reported previously from our group. Mitral regurgitation (MR) frequently accompanies aortic valve disease that at times may also require repair. Totally endoscopic surgery in such cases has not been tested. We present a surgical technique for a totally endoscopic approach to aortic valve replacement and concomitant mitral valve repair for primary and secondary MR. An aortotomy incision was used avoiding an atriotomy, which results in an increase in cross-clamp (XC) and cardiopulmonary bypass (CPB) times that could be associated with higher mortality and morbidity. Neochords (artificial chordae tendineae) were used for primary MR and an edge-to-edge approach for secondary MR. TEAVR and concomitant mitral valve repair can be performed successfully with reasonable XC and CPB times with excellent short-term results.


Transcriptomic analysis using dual RNA sequencing revealed a Pathogen-Host interaction after Edwardsiella anguillarum infection in European eel (Anguilla anguilla).
Many studies have explored differentially expressed genes (DEGs) between some pathogens and hosts, but no study has focused on the interaction of DEGs between Edwardsiella anguillarum (Ea) and Anguilla anguilla (Aa). In this study, we examined the interactions of DEGs during Ea infection and Aa anti-infection processes by dual RNA sequencing. Total RNA from in vitro and in vivo (Aa liver) Ea culture was extracted. Using high-throughput transcriptomics, significant DEGs that were expressed between Ea cultured in vitro versus in vivo and those in the liver of the infected group versus control group were identified. Protein-protein interactions between the pathogen and host were explored using Cytoscape according to the HPIDB 3.0 interaction transcription database. The results showed that the liver in the infection group presented with severe bleeding and a large number of thrombi in the hepatic vessels. We found 490 upregulated and 398 downregulated DEGs of Ea in vivo versus Ea cultured in vitro, and 2177 upregulated and 970 downregulated genes in the liver of the infected eels. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the pathogen DEGs revealed that the upregulated genes were mainly enriched in migration, colonization, biofilm formation, and significantly enriched in ABC transport and quorum sensing; the downregulated genes were mainly involved in metabolism, information transduction, organelle formation, enzyme catalysis, molecular transport, and binding. GO of the host DEGs showed that metabolic process, catalytic activity, single organism metabolic process, small molecule binding, nucleotide binding, nucleotide phosphate binding, and anion binding were markedly enriched. Finally, we found that 79 Ea and 148 Aa proteins encoded by these DEGs were involved in an interaction network, and some pathogen (DegP, gcvP, infC, carB, rpoC, trpD, sthA, and FhuB) and host proteins (MANBA, STAT1, ETS2, ZEP1, TKT1, NMI and RBPMS) appear to play crucial roles in infection. Thus, determining the interaction networks revealed crucial molecular mechanisms underlying the process of pathogenic infection and host anti-infection.


Lipid signaling in experimental epilepsy.
Glutamate release activates signaling pathways important for learning and memory, and over-stimulation of these pathways during seizures leads to aberrant synaptic plasticity associated with hyper-excitable, seizure-prone states. Seizures induce rapid accumulation of membrane lipid-derived fatty acids at the synapses which, evidence suggests, regulate maladaptive connectivity. Here we give an overview of the significance of the arachidonyl- and inositol-derived messengers, prostaglandins (PGs) and diacylglycerol (DAG), in experimental models of epilepsy. We use studies conducted in our own laboratory to highlight the pro-epileptogenic role of cyclooxygenase-2 (COX-2) and its products, the PGs, and we discuss the possible mechanisms by which PGs may regulate membrane excitability and synaptic transmission at the cellular level. We conclude with a discussion of AA-DAG signaling in synaptic plasticity and seizure susceptibility with an emphasis on recent studies in our laboratory involving DAG kinase epsilon (DGKepsilon)-knockout mice.


The regulatory peptide system of the large bowel in equine grass sickness.
In recent years, distinct changes in regulatory peptides have been found in a number of gastrointestinal diseases. Grass sickness is a fatal disease of horses for which the etiology has yet to be fully ascertained. In this study, the peptide-containing nerves and ganglionic and mucosal endocrine cells of the ileum, colon and rectum were investigated in horses with sub-acute or chronic grass sickness and compared with normal controls using immunocytochemistry, at both the light and electron microscopical levels, and radioimmunoassay. A substantial loss of both peptide-containing cells and nerves was found in all of the sick horses, particularly in the ileum. Electron microscopy revealed marked degeneration of nerves in the gut wall. Fibers containing granules immunostained for substance P or VIP, using the immunogold staining technique, underwent extensive degranulation in grass sickness, with the formation of multiple vacuoles. Radioimmunoassay of peptide content also showed that the most drastic changes occurred in the ileum. For example, VIP content was significantly reduced from 109 +/- 19.8 (mean +/- SEM) pmoles/g in controls to 6.8 +/- 1.4 pmoles/g in grass sickness (p less than 0.001) and substance P from 65.9 +/- 8.1 to 31.3 +/- 9.5 (p less than 0.02). These results may have applications in the diagnosis and treatment of grass sickness.


Modulation of human breast cancer cell adhesion by estrogens and antiestrogens.
In order to study the effect of estrogens and antiestrogens on the adhesive properties of human breast cancer cells, the attachment on endothelial cells (EC), on subendothelial extracellular matrix (ECM) and on ECM components (collagen I and IV, laminin, fibronectin) of estrogen-dependent (MCF-7, ZR75-1) and estrogen-independent (BT-20) breast cancer cell lines was investigated. The cells were grown under conditions of controlled exposure to estrogen [17 beta-estradiol (E2)] and/or antiestrogens [tamoxifen (Tam) or 4-hydroxytamoxifen (OH-Tam)]. Treatment by E2 enhanced the ability of ZR75-1 cells to adhere to the various substrates, which contrasts with the observed absence of effects with the BT-20 cells. Similarly, Tam or OH-Tam induced a reduction of the adhesion of ZR75-1 tumor cell, but not of BT-20 cells. This effect was reversed by competing concentrations of E2. The effects on MCF-7 cell adhesion were similar to those described for ZR75-1 cells, but could not be reproducibly observed. Adhesion assays carried out with ZR75-1 cells grown in the absence or presence of phenol red, a pH indicator which behaves as a weak estrogen, led to a similar pattern of cell attachment. Conditioned media harvested from E2- or Tam-treated ZR75-1 cells failed to induce any effect on adhesion of other ZR75-1 cells grown in E2-deprived medium, suggesting that secretory activities are not required for the control of cell adhesiveness. The results suggest that estrogens and antiestrogens can control the adhesive behavior of breast tumor cells through their hormone responsive structures possibly by regulating expression of cell adhesion proteins and/or their cell surface receptors.


Hereditary coproporphyria: comparison of molecular and biochemical investigations in a large family.
Hereditary coproporphyria (HCP) is the least common of the three autosomal dominant acute porphyrias. To compare the sensitivity of metabolite measurements for the identification of asymptomatic HCP, we carried out a molecular and biochemical investigation of a large family in which HCP is caused by a previously unreported frameshift mutation (c.119delA). Thirteen of 19 asymptomatic family members, aged 10-72 years, were shown by mutational analysis to have HCP. The faecal coproporphyrin isomer III:I ratio was increased in all of these 13 family members; faecal total porphyrin concentration and urinary porphyrin excretion were increased in 11 and 8 of them, respectively. Plasma porphyrin concentrations were marginally increased in three individuals and plasma fluorescence emission scanning showed a porphyrin peak at 618 nm in two of these. Our results add to the evidence that an increased faecal porphyrin coproporphyrin III:I ratio is a highly sensitive test for the detection of clinically latent HCP in individuals over the age of 10 years; its sensitivity below this age remains uncertain. They also show that plasma fluorescence emission scanning is not useful for the investigation of families with HCP.


Outcomes of bilateral carpal tunnel syndrome treatment - A systematic review and meta-analysis.
Carpal tunnel syndrome (CTS) is the predominant compressive neuropathy among adults worldwide. However, evidence regarding treatment alternatives and their outcomes, especially with bilateral carpus involvement, is inconclusive. To analyze the clinical and surgical outcomes of bilateral CTS treatment using the visual analog scale (VAS) and Boston questionnaire, a systematic review was conducted according to PRISMA guidelines. After evaluating 129 articles from different databases, nine papers with low bias risk were included in the study. These studies were assessed for methodological quality, both in scale and degree, ensuring bias identification and independence of data extraction. Eligible articles were those in Portuguese, Spanish, and English, with no publication time limit. The outcomes assessed were the standardized mean differences (SMDs) on the symptom severity scale (SSS), functional state scale (FSS), and VAS. In the clinical treatment group, a positive effect was observed on the SSS (SMD: 0.53), FSS (SMD: 0.47), and VAS (SMD decrease: 2.52) at the one-month follow-up. In the surgical-treatment group, a positive effect was observed on the SSS (SMDs: 1.97 for endoscopic and 1.55 for open surgery), FSS (SMDs: 1.52 for endoscopic and 1.77 for open surgery), and VAS (SMDs: 2.19 for endoscopic and 2.59 for open surgery) at the one-month follow-up. Significant improvements in VAS, SSS, and FSS were observed at the three-month follow-up. Current evidence in both clinical and surgical treatments demonstrates their effectiveness, as they potentially improve symptom severity, functional status, and pain intensity in patients with bilateral CTS during one- and three-month follow-up periods.


Pulse contour analysis versus thermodilution in cardiac surgery patients.
Previous studies have demonstrated that there is a lack of agreement between intermittent cold bolus thermodilution (ICO) and a semicontinuous method with dilution of heat (CCO) in cardiac surgical patients following hypothermic extracorporeal circulation (HCPB). Therefore, the aim of the present study was to compare both ICO and CCO with continuous pulse contour analysis (PCCO): a method based on a fundamentally different principle of determining cardiac output (CO). A prospective criterion standard study of 25 cardiac surgery patients undergoing HCPB. Cardiac output was determined using the three methods (ICO, CCO, and PCCO) before and after HCPB up to 12 h after arrival on the ICU. Bias and precision were evaluated. A total of 380 triple determinations of CO could be analyzed. During the entire study period bias PCCO-ICO was -0.14 l*/min (precision 1.16 l*/min) and bias CCO-ICO was -0.40 l*/min (precision 1.25 l*/min). Up to 45 min after bypass PCCO agreed with ICO (bias -0.21 l*/min, precision 1.37 l*/min), while bias CCO-ICO was -1.30 l*/min (precision 1.45 l*/min). The agreement between PCCO and ICO in contrast to CCO in the first 45 min after HCPB indicates that CCO underestimates CO during this period.


Hypersensitivity reactions to anti-SARS-CoV-2 vaccines: Basophil reactivity to excipients.
Basophil activation test (BAT) can tackle multiple mechanisms underlying acute and delayed hypersensitivity to drugs and vaccines and might complement conventional allergy diagnostics but its role in anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine-related hypersensitivity is ill-defined. Therefore, 89 patients with possible hypersensitivity (56 % with delayed mucocutaneous manifestations) to anti-SARS-CoV-2 vaccines were tested with BAT for Macrogol 3350, DMG-PEG 2000, PEG 20000, polysorbate-80 and trometamol and compared to 156 subjects undergoing pre-vaccine BAT. A positive BAT was associated with delayed reaction onset (p = 0.010) and resolution (p = 0.011). BAT was more frequently positive to DMG-PEG 2000 than to other excipients in both groups (p < 0.001). DMG-PEG 2000 reactivity was less frequent in vaccine-naïve (6 %) than vaccinated subjects (35 %, p < 0.001) and associated with mRNA-1273 vaccination. DMG-PEG 2000 BAT might therefore have a diagnostic role in subjects with delayed hypersensitivity reactions. Natural immunity might be a key player in basophil activation.


Discrimination by parity is a prerequisite for assessing induction of labour outcome - cross-sectional study.
To demonstrate that studies on induction of labour should be analyzed by parity as there is a significant difference in the labour outcome among induced nulliparous and multiparous women. Obstetric outcome, specifically caesarean section rates, among induced term nulliparous and multiparous women without a previous caesarean section were analyzed in this cross-sectional study using the Robson 10 group classification for the year 2016. In the total number of 8851 women delivered in 2016, the caesarean section rates among nulliparous women in spontaneous and induced labour, Robson groups 1 and 2A, were 7.84% (151/1925) and 32.63% (437/1339) respectively and among multiparous (excluding those women with a previous caesarean section), Robson group 3 and 4A were 1%(24/2389) and 4.37% (44/1005), respectively. Pre labour caesarean rates for nulliparous and multiparous women, Robson groups 2B and 4B (Robson M, Fetal Matern Med Rev, 12; 23-39, 2001) were 3.91% (133/3397) and 2.86% (100/3494), of the respective single cephalic cohort at term. The data suggests that studies on induction of labour should be analyzed by parity as there is a significant difference between nulliparous and multiparous women.


Cone-beam computed tomography for routine orthodontic treatment planning: a radiation dose evaluation.
Because of the advantages and possibilities of cone-beam computed tomography (CBCT), orthodontists use this method routinely for patient assessment. The aim of this study was to compare the radiation doses for conventional panoramic and cephalometric imaging with the doses for 2 different CBCT units and a multi-slice CT unit in orthodontic practice. The absorbed organ doses were measured by using an anthropomorphic phantom loaded with thermoluminescent dosimeters at 16 sites related to sensitive organs. The 4 devices (Sirona DS Plus [Sirona Dental Systems, Bernsheim, Germany], i-CAT [Imaging Sciences International, Hatfield, Pa], NewTom DVT 9000 [QR, Verona, Italy], and Somatom Sensation [Siemens Medical Solutions, Erlangen, Germany]) were used with standard protocols and, when possible, in the auto-exposure mode. Equivalent and effective doses were calculated. The calculation of the effective doses was based on the International Commission on Radiological Protection's 2005 recommendations. The lowest organ dose (13.1 microSv) was received by the thyroid gland during conventional panoramic and lateral cephalometric imaging. The highest mean organ dose (15,837.2 microSv) was received by the neck skin from the multi-slice CT. The effective dose was also lower for the panoramic and lateral cephalometric device (10.4 microSv), and highest for the multi-slice CT (429.7 microSv). From a radiation-protection point of view, conventional images still deliver the lowest doses to patients. When 3-dimensional imaging is required in orthodontic practice, a CBCT should be preferred over a CT image. Further studies are necessary to justify the routine use of CBCT in orthodontic treatment planning.


A convenient preparation of a disaccharide motif and its role in the cytotoxicity of the triterpenoid saponin, alpha-hederin.
The sugar structures of triterpenoid saponins, such as alpha-hederin, are intimately associated with their antitumor activities and other biological activities. The alpha-L: -rhamnopyranosyl-(1-->2)-alpha-L: -arabinopyranoside group of alpha-hederin alters the cytotoxicity of its aglycon, hederagenin. This study explored the role of this saccharide unit in the cytotoxic effect of alpha-hederin and the possibility of its use as a carrier moiety in prodrugs of anticancer agents. A new convenient and practical procedure for the preparation of 4-methoxybenzoyl-2,3,4-tri-O-benzoyl-alpha-L: -rhamnopyranosyl-(1-->2)-3,4-O-dibenzoyl-beta-L: -arabinopyranoside (2) from 4-methoxybenzoyl-beta-L: -arabinopyranoside was accomplished using four steps with an overall yield of 63%. The use of BF(3)-OEt(2) as a catalyst in the glycosylation step in this procedure had a large advantage over the TMSOTf catalyst used in the usual method. Moreover, the key intermediate obtained in this procedure, 4-methoxybenzoyl-2,3,4-tri-O-benzoyl-alpha-L: -rhamnopyranosyl-(1-->2)-alpha-L: -arabinopyranoside (7), was selectively transformed to 4-methoxybenzoyl-2,3,4-tri-O-benzoyl-alpha-L: -rhamnopyranosyl-(1-->2)-4-O-acetyl-alpha-L: -arabinopyranoside (9) and 4-methoxybenzoyl-2,3,4-tri-O-benzoyl-alpha-L: -rhamnopyranosyl-(1-->2)-3-O-benzoyl-beta-L: -arabinopyranoside (10). These derivatives did not show any cytotoxicity against human cancer cell lines. Thus the 3-O-alpha-L: -rhamnopyranosyl-(1-->2)-alpha-L: -arabinopyranoside could be used as a nontoxic carrier moiety to enhance the activity of anticancer drugs.


Vectors and malaria transmission in deforested, rural communities in north-central Vietnam.
Malaria is still prevalent in rural communities of central Vietnam even though, due to deforestation, the primary vector Anopheles dirus is uncommon. In these situations little is known about the secondary vectors which are responsible for maintaining transmission. Basic information on the identification of the species in these rural communities is required so that transmission parameters, such as ecology, behaviour and vectorial status can be assigned to the appropriate species. In two rural villages--Khe Ngang and Hang Chuon--in Truong Xuan Commune, Quang Binh Province, north central Vietnam, a series of longitudinal entomological surveys were conducted during the wet and dry seasons from 2003-2007. In these surveys anopheline mosquitoes were collected in human landing catches, paired human and animal bait collections, and from larval surveys. Specimens belonging to species complexes were identified by PCR and sequence analysis, incrimination of vectors was by detection of circumsporozoite protein using an enzyme-linked immunosorbent assay. Over 80% of the anopheline fauna was made up of Anopheles sinensis, Anopheles aconitus, Anopheles harrisoni, Anopheles maculatus, Anopheles sawadwongporni, and Anopheles philippinensis. PCR and sequence analysis resolved identification issues in the Funestus Group, Maculatus Group, Hyrcanus Group and Dirus Complex. Most species were zoophilic and while all species could be collected biting humans significantly higher densities were attracted to cattle and buffalo. Anopheles dirus was the most anthropophilic species but was uncommon making up only 1.24% of all anophelines collected. Anopheles sinensis, An. aconitus, An. harrisoni, An. maculatus, An. sawadwongporni, Anopheles peditaeniatus and An. philippinensis were all found positive for circumsporozoite protein. Heterogeneity in oviposition site preference between species enabled vector densities to be high in both the wet and dry seasons allowing for year round transmission. In rural communities in north central Vietnam, malaria transmission was maintained by a number of anopheline species which though collected feeding on humans were predominantly zoophilic, this behaviour allows for low level but persistent malaria transmission. The important animal baits--cattle and buffalo--were kept in the village and barrier spraying around these animals may be more effective at reducing vector densities and longevity than the currently used indoor residual spraying.


Anti-tumor angiogenesis effect of a new compound: B-9-3 through interference with VEGFR2 signaling.
B-9-3, a derivative of harmine, was first synthesized in our laboratory. We have reported that B-9-3 has an anti-proliferative effect against human lung cancer cells via induction of apoptosis and inhibition of cell migration. In the present study, we first studied that the anti-tumor angiogenesis effect and the molecular mechanism of B-9-3-induced tumor vascular degrade and mortify in lung cancer. In vitro, the results showed that B-9-3 selectively inhibited the proliferation of endothelial cells IC50 = 6.16 μg/ml) and vascular fibroblasts (IC50 = 12.59 μg/ml) and induced regression of tumor cells of the following: Lewis lung carcinoma (LLC), Mouse fore-stomach carcinoma (MFC), Human ovarian cancer (SK-OV-3), and prostate cancer (22RV1). Moreover, B-9-3 could significantly increase the apoptosis rate (80.95 %) of vascular endothelial cells, while inhibiting migration of endothelial cells, capillary tube formation of endothelial cells, neovascularization of the rat thoracic aorta ring, and the angiogenesis of chick chorioallantoic membrane (CAM) predominantly through blocking VEGFR2 signaling pathway. In vivo, we investigated the anti-tumor rate and the signal transduction mechanism of B-9-3 by LCC-bearing C57BL/6 mice. The data showed that the tumor inhibition ratio of high dose (20 mg/kg) of B-9-3 was 72.9 %, and quantification of CD34 marker indicated a marked reduction in the number of neovessels after B-9-3 treatment as compared with control group (66.87 %). Remarkably, using IHC and q-RT-PCR, we found that downregulation of the expression of VEGFR2, VEGF-A, and TGFβ1 in tumor confers enhancement to the angiogenesis effect of B-9-3. These data suggest that the angiogenesis inhibitor B-9-3 selectively induces apoptosis of endothelial cells, in part through disruption of VEGF-A/VEGFR2 signaling.


Birth defects in a national cohort of pregnant women with HIV infection in Italy, 2001-2011.
We used data from a national study of pregnant women with HIV to evaluate the prevalence of congenital abnormalities in newborns from women with HIV infection. Observational study. University and hospital clinics. Pregnant women with HIV exposed to antiretroviral treatment at any time during pregnancy. The total prevalence of birth defects was assessed on live births, stillbirths, and elective terminations for fetal anomaly. The associations between potentially predictive variables and the occurrence of birth defects were expressed as odds ratios (ORs) with 95% confidence intervals (95% CIs) for exposed versus unexposed cases, calculated in univariate and multivariate logistic regression analyses. Birth defects, defined according to the Antiretroviral Pregnancy Registry criteria. A total of 1257 pregnancies with exposure at any time to antiretroviral therapy were evaluated. Forty-two cases with major defects were observed. The total prevalence was 3.2% (95% CI 1.9-4.5) for exposure to any antiretroviral drug during the first trimester (23 cases with defects) and 3.4% (95% CI 1.9-4.9) for no antiretroviral exposure during the first trimester (19 cases). No associations were found between major birth defects and first-trimester exposure to any antiretroviral treatment (OR 0.94, 95% CI 0.51-1.75), main drug classes (nucleoside reverse transcriptase inhibitors, OR 0.95, 95% CI 0.51-1.76; non-nucleoside reverse transcriptase inhibitors, OR 1.20, 95% CI 0.56-2.55; protease inhibitors, OR 0.92, 95% CI 0.43-1.95), and individual drugs, including efavirenz (prevalence for efavirenz, 2.5%). This study adds further support to the assumption that first-trimester exposure to antiretroviral treatment does not increase the risk of congenital abnormalities.


Estimating Microplastics related to Laundry Wash and Personal Care Products released to Wastewater in Major Estonian Cities: a comparison of calculated and measured microplastics.
Microplastics (MPs) research still at the budding stage in Estonia. A theoretical model build on substance flow analysis principles was developed. The goal of this study is to broaden understanding of MPs-types in wastewater and their contribution from known sources, quantify their presence based on model prediction and in-situ measurements. The authors estimate MPs from laundry wash (LW) and personal care products (PCPs)) in wastewater in Estonia. We found out that total estimated MPs load per capita from PCPs and LW in Estonia were between 4.25 - 12 tons/year, 3.52 - 11.24 tons / year respectively, and estimated load ended up in wastewater were between 700 - 30,000 kg/yr. and 2 - 1500 kg/yr. in WWTPs influent and effluent stream respectively. Finally. We conducted a comparison between estimated MPs load and on-site sample analysis and observed a medium-high level of MPs being discharged into the environment annually. During quantification and chemical characterization using µFTIR analysis, we found that microfibers with a length of 0.2-0.6 mm accounted for over 75% of the total MPs load in the effluent samples collected from four coastal WWTPs in Estonia. The estimation avails us broader overview about the theoretical MPs load in wastewater and gain valuable insight into developing process methods that prevent MPs accumulation in sewage sludge for safe application in agriculture.


Should less-invasive aortic valve replacement be avoided in patients with pulmonary dysfunction?
In patients with pulmonary dysfunction, it is unclear whether a less-invasive approach for aortic valve replacement is well tolerated or even beneficial. We investigated whether a partial upper J-incision for aortic valve replacement leads to more favorable outcomes than a full sternotomy in patients with chronic lung disease by using forced expiratory volume in 1 second as a surrogate. From January 1995 to July 2010, 6931 patients underwent primary isolated aortic valve replacement; 655 had forced expiratory volume in 1 second measured and expressed as percent of predicted (FEV1%; 368 via J-incision, 287 via full sternotomy). Postoperative outcomes were compared among 223 propensity-matched pairs. Patients diagnosed with chronic lung disease had longer median intensive care unit (41 vs 27 hours, P = .001) and postoperative (7.1 vs 6.1 days, P < .0001) lengths of stay than those without chronic lung disease. At normal values of FEV1%, little difference was observed in either of these times for J-incision versus full sternotomy; however, at progressively lower FEV1%, these times lengthened, with increasing benefit for J-incision. Among propensity-matched patients, other postoperative complications were similar. Early survival (93% vs 89% at 1 year, P = .07) was possibly higher in matched patients with J-incision, but late survival was similar (P = .9). Patients with FEV1% less than 50 who underwent J-incision had the greatest survival advantage, which persisted for 5 years. In patients with preoperative respiratory dysfunction, a less-invasive partial upper J-incision for aortic valve replacement can lead to more favorable outcomes than a full sternotomy, including shorter intensive care unit and postoperative lengths of stay and better early survival, which are amplified with decreasing pulmonary function.


Rapid identification of fungi from respiratory samples by Bruker Biotyper matrix-assisted laser desorption/ionisation time-of-flight using ID-FUNGI plates.
Identification of moulds is crucial for the clinical management of patients. The goal of this study was to evaluate the new ID-FUNGI plate (IDFP) for the identification of moulds by MALDI Biotyper. IDFP was compared with Sabouraud with gentamicin and chloramphenicol plate (SAB) for the identification of 80 moulds from respiratory samples and eight reference strains. With the direct transfer method, species identification rose from 6% with SAB to 68% with IDFP using score cut-off 2 and from 20 to 75% using cut-off 1.7 (p < 0.001). Our study highlights that the new IDFP improves mycological diagnostic and workflow in laboratories.


Design and synthesis of antiangiogenic/heparin-binding arginine dendrimer mimicking the surface of endostatin.
We designed and synthesized the antiangiogenic arginine-rich dendrimers, TX-1943 and TX-1944, which mimic the surface structure of endostatin. TX-1944 containing 16 arginine residues is more similar in surface structure to endostatin than TX-1943 with eight arginine residues, and has stronger in vivo antiangiogenic activity at 10 microg/CAM in the chicken embryo chorioallantoic membrane (CAM) assay. TX-1944 also has stronger activity to bind heparin and to inhibit the growth of rat lung endothelial (RLE) cells than TX-1943.


Grafting of polymers via ring-opening polymerization for electrochemical assay of alkaline phosphatase activity.
As an important hydrolytic enzyme, abnormal activity of alkaline phosphatase (ALP) is closely associated with a variety of diseases. It has been identified as an important diagnostic indicator for clinical hepatobiliary and bone diseases. Herein, a novel electrochemical sensor based on signal amplification strategy through ring-opening polymerization (ROP) has been developed to assay of ALP activity. First of all, 3-mercaptopropanoic acid (MPA) was employed as a cross-linking agent to attach O-phosphoethanolamine to the electrode surface via amide bond. Then, ALP catalyzed the hydrolysis of phosphate monoester structures to hydroxyl groups, which could initiate ROP reaction. The polymer grafted on the electrode surface contains a large number of ferrocene electroactive molecules, which effectively increased the signal output of the electrochemical sensor and improved the sensitivity of ALP activity detection. Under optimum conditions, this electrochemical sensor rendered a satisfactory linear dependence over the range from 20 to 120 mU mL-1, with a low detection limit of 0.66 mU mL-1. Furthermore, this strategy presented satisfactory selectivity and interference resistance in human serum sample, and compared with clinical data, the relative error of the results obtained by this method was less than 5%. Thus, this method showed considerable potential for the detection of ALP activity in clinical application.


Retrobulbar cellulitis and abscessation: focus on short- and long-term concurrent ophthalmic diseases in 41 dogs.
To describe concurrent ophthalmic diseases in dogs with retrobulbar cellulitis and abscessation. Retrospective analysis of clinical records of dogs with retrobulbar inflammation. Forty-one dogs were diagnosed with retrobulbar inflammation; of these, 23 presented with abscessation and two with zygomatic sialoadenitis. Diagnosis was based on orbital ultrasound, MRI, CT and cytological and microbiological examination of fluid or tissue samples. Management involved evacuation of fluid contents using ultrasound-guided fine-needle aspiration via the pterygopalatine fossa or orbitotomy. Patients received systemic antibiotics (except for one with sialoadenitis), glucocorticoids, non-steroidal anti-inflammatory drugs, opioids and fluid therapy. At initial presentation one or more ophthalmic complications were reported in 19 dogs (46%) including internal ophthalmoplegia (n=5), blindness due to optic nerve damage (n=5), facial nerve paralysis (n=3), prolapse of the third eyelid gland (n=3), corneal ulceration (n=8), anterior uveitis (n=4), chorioretinitis (n=3), retinal detachment (n=2) and increased intraocular pressure (n=7). Information on ophthalmic complications after cessation of active inflammation was available for 33 patients. One or more concurrent disease was found in 10 cases (30%): in addition to persistent neurological deficits and third eyelid gland prolapse reported at initial presentation, visual deficits after retinal re-attachment (n=2), loss of corneal sensation (n=1), corneal oedema (n=1), corneal fibrosis (n=4), corneal lipidosis (n=1) and strabismus after suspected fibrosis (n=2) were diagnosed. Ophthalmic complications are common in patients with retrobulbar inflammation indicating that these patients should undergo ophthalmic assessment and follow-up.


Combined therapy for small cell undifferentiated carcinoma of the lung.
Fifty-eight patients with small cell lung cancer were treated from September, 1974, to March, 1976, with combined chemotherapy and radiotherapy. Surgical resection of the lung lesion was performed in three patients, and a number of surgical diagnostic methods were carried out in the remaining patients with unresectable of disseminated lesions. Nineteen patients were from the Veterans Administration Hospital and 39 from Indiana University Medical Center. The median Karnofsky performance status was 60. Thirty-nine patients had extensive disease, and 19 had disease limited to the chest and supraclavicular area. All patients received chest radiotherapy and prophylactic whole brain radiation. Adriamycin, cyclophosphamide, and vincristine were given on day 1 and continued every 3 weeks. There were 27 (48 percent) partial remissions of a median duration of 26 weeks. There were 25 patients (43 percent) with complete remission. The median survival for the entire group was 51 weeks. Six of 58 patients (10 percent) are alive and disease free from 24 to 38 months after treatment. Six of 19 patients with limited disease (32 percent) are presently alive and disease free. This includes one patient in whom surgical resection was performed. Combined therapy influences favorably the prognosis of small cell cancer of the ling, expecially in those patients with limited disease and favorable performance status.


PACAP signaling in stress: insights from the chromaffin cell.
Pituitary adenylate cyclase-activating polypeptide (PACAP) was first identified in hypothalamus, based on its ability to elevate cyclic AMP in the anterior pituitary. PACAP has been identified as the adrenomedullary neurotransmitter in stress through a combination of ex vivo, in vivo, and in cellula experiments over the past two decades. PACAP causes catecholamine secretion, and activation of catecholamine biosynthetic enzymes, during episodes of stress in mammals. Features of PACAP signaling allowing stress transduction at the splanchnicoadrenomedullary synapse have yielded insights into the contrasting roles of acetylcholine's and PACAP's actions as first messengers at the chromaffin cell, via differential release at low and high rates of splanchnic nerve firing, and differential signaling pathway engagement leading to catecholamine secretion and chromaffin cell gene transcription. Secretion stimulated by PACAP, via calcium influx independent of action potential generation, is under active investigation in several laboratories both at the chromaffin cell and within autonomic ganglia of both the parasympathetic and sympathetic nervous systems. PACAP is a neurotransmitter important in stress transduction in the central nervous system as well, and is found at stress-transduction nuclei in brain including the paraventricular nucleus of hypothalamus, the amygdala and extended amygdalar nuclei, and the prefrontal cortex. The current status of PACAP as a master regulator of stress signaling in the nervous system derives fundamentally from the establishment of its role as the splanchnicoadrenomedullary transmitter in stress. Experimental elucidation of PACAP action at this synapse remains at the forefront of understanding PACAP's role in stress signaling throughout the nervous system.


Conservative management of mesh-site infection in hernia repair surgery: a case series.
The aim of this study is to assess the outcome of conservative management of infected mesh grafts following abdominal wall hernia repair. This study retrospectively examined the charts of patients who developed mesh-site infection following surgery for abdominal hernia repair to determine how effective conservative management in the form of antibiotics and wound management was on the resolution of infection and wound healing. Over a period of 30 months, 13 patients developed infected mesh grafts post-hernia repair surgery. Twelve patients were successfully treated conservatively with local wound care and antibiotics if clinically indicated. One patient returned to theatre to have the infected mesh removed. Of the patients that healed eleven were treated with negative pressure wound therapy (VAC(®)). This series of case studies indicate that conservative management of abdominal wall-infected hernia mesh cases is likely to be successful.


A specific binding protein for the moulting hormone ecdysterone in locust haemolymph.
Specific binding of 3H-ecdysterone to a high mol. wt protein from Locusta migratoria haemolymph was shown by gel filtration. The hormone-protein complex shows a dissociation constant Kd congruent to 3.10(-7) M, and the concentration of binding sites varies during the last larval instar.


H3K27 acetylation activated-CCS regulates autophagy and apoptosis of lung cancer by alleviating oxidative stress.
Copper chaperone for superoxide dismutase (CCS) is abnormally expressed in various human malignancies. However, the function and mechanism of CCS in lung cancer progression remain unclear. In the current study, CCS protein and mRNA levels were found to be increased in lung adenocarcinoma (LUAD) tissue and cell lines. Patients with higher CCS levels had a poorer prognosis. Decreasing the enrichment of histone H3 Lys27 acetylation (H3K27ac) by A-485 inhibited CCS expression. CCS depletion upregulated reactive oxygen species (ROS) levels, aggravated oxidative stress, inhibited autophagy, inhibited cell survival, and promoted apoptosis. The treatment of antioxidant N-Acetyl-L-cysteine (NAC) rescued these changes induced by CCS depletion. CCS also was found to be related to the immune infiltration of CD8 + T cells and regulatory T cells in LUAD. These data indicated that overexpression of CCS activated by H3K27 acetylation relieved oxidative stress, promoted autophagy, and inhibited apoptosis. CCS may be regarded as a potential therapeutic target for LUAD.


Histochemistry of glycoconjugates in the gallbladder epithelium of ten animal species.
A battery of seven lectins and several conventional mucin histochemical techniques were used to identify the epithelial mucins of the gallbladder of ten species: man, rabbit (Oryctolagus cuniculus, mammalia), hamster (Mesocricetus auratus, mammalia), chicken (Gallus gallus, bird), sparrow (Passer domesticus, bird), moorish gecko (Tarentola mauritanica, reptilia), ladder snake (Elaphe scalaris, reptilia), lake frog (Rana perezi, amphibia), natterjack toad (Bufo calamita, amphibia) and gilthead sea bream (Sparus auratus, fish). Glycogen was found in the epithelial lining of the reptilian and amphibian gallbladders. Sulphate and carboxyl groups were frequently found in the same species, except in the ladder snake and natterjack toad gallbladders where only sulphate groups were identified. Sialic acid residues were detected in man, rabbit, bird, T. mauritanica, R. perezi and fish gallbladders. ConA binding pattern was similar in the ten species studied. In the human gallbladder only PNA failed to label the luminal surface, while the glands were only unreactive to DBA. The human gallbladder showed a large variety of saccharides. The present results suggest that no relation exists between the composition of the gallbladder mucins and the situation of the species in the phylogenetic scale.


Synthesis and secretion of corticotropins, melanotropins, and endorphins by rat intermediate pituitary cells.
The synthesis and secretion of various intermediate pituitary proteins was studied by using dispersed intermediate pituitary cell suspensions. Control studies indicated that the isolated cells were obtained in good yield and that after more than 24 h in culture the isolated cells continued to synthesize a collection of proteins similar to those found in freshly extracted intermediate pituitary tissue. Rat intermediate pituitary cells synthesized a molecule (Mr = 30,000; called 30K) that contained antigenic determinants for beta-endorphin, gamma-lipotropin, corticotropin (ACTH), and 16K fragment (the NH2-terminal region of mouse tumor cell pro-ACTH/endorphin). This 30K molecule, two high molecular weight forms of ACTH(13K and 20K), and 16K fragment were all shown to be glycoproteins. Continuous labeling and pulse-chase incubations were used to define the intracellular biosynthetic processing of the 30K molecule. After a 15-min pulse incubation the 30K molecule was the only labeled protein containing antigenic determinants for beta-endorphin, gamma-lipotropin, ACTH, or 16K fragment. A beta-lipotropin-like molecule served as a biosynthetic intermediate in the production of proteins similar to beta-endorphin and gamma-lipotropin. Methionine-enkephalin and alpha-endorphin were not major products in the intermediate lobe cells. Molecules similar to alpha-melanocyte-stimulating hormone and corticotropin-like intermediate lobe peptide (ACTH(18-39)) were also derived from the same 30K molecule; 20K ACTH served as a biosynthetic intermediate in this conversion. In rat intermediate pituitary cells ACTH(1-39) was not a major final product of the intracellular biosynthetic processing of the 30K molecule. The 30K molecule also served as a precursor to a protein similar to mouse tumor cell 16K fragment and related smaller proteins. With rat intermediate pituitary cells, pulse-chase experiments utilizing [35S]methionine demonstrated almost quantitative conversion of the 30K precursor into labeled proteins similar to beta-endorphin and alpha-melanocyte-stimulating hormone. In the absence of added secretagogues, small amounts of all of the smaller proteins derived from the 30K precursor were secreted coordinately into the culture medium.


Exuberant Biofilm infection in a lateral canal as the cause of short-term endodontic treatment failure: report of a case.
This article describes a case in which signs and symptoms persisted in spite of the endodontic treatment following high standards to characterize a short-term failure. After several appointments of root canal treatment in an attempt to resolve persistent symptoms, including 106 days of calcium hydroxide intracanal medication, periradicular surgery was performed, and the root apex and the lesion were subjected to histologic and histobacteriologic analyses. The lesion was diagnosed as a cyst, and the main root canal was free of bacteria and debris. The cause of the short-term failure was an exuberant bacterial biofilm colonizing a lateral canal in the apical root segment. This case report highlights one of the major problems of modern endodontic therapy; bacteria located in areas distant from the main root canal can remain unaffected by treatment procedures and maintain disease. The challenge for researchers and clinicians that arises from this problem is to develop strategies, instruments, or substances that can reach those areas and achieve sufficient reduction in the infectious bioburden to permit predictable periradicular healing.


Expression of Concern.
Expression of Concern: 'miR-377-3p drives malignancy characteristics via upregulating GSK-3β expression and activating NF-κB pathway in hCRC cells', by Wei-Ying Liu, Zhen Yang, Qi Sun, Xi Yang, Yang Hu, Hong Xie, Hui-Jie Gao, Li-Ming Guo, Jian-Ying Yi, Min Liu, and Hua Tang (2017); J Cell Biochem. 2018; 2124-2134: This Expression of Concern is for the above article, published online on 31 August 2017, in Wiley Online Library (https://doi.org/10.1002/jcb.26374), and has been published by agreement between the the journal's Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. Concerns about data integrity were raised by an independent source regarding a possible duplication in Figure 4D. The authors admit an error at compilation of the figures and provided a correction. After a detailed investigation and analysis of the raw data, concerns remain regarding the integrity of the data and the conclusions of the article. The authors' institution, Tianjin Medical University, has been contacted, and an investigation of this paper from the group of Hua Tang is underway. While the conclusions of this investigation are awaited, the journal has decided to issue this Expression of Concern.


Composition gradient static light scattering: a new technique for rapid detection and quantitative characterization of reversible macromolecular hetero-associations in solution.
A method for rapid detection and quantitative characterization of associations between two proteins in solution is presented, based upon continuous measurement of the Rayleigh light scattering and UV absorbance of a solution whose composition is varied with time in a controlled fashion. Collection and subsequent analysis of data are both rapid and semiautomatic. The results of validation experiments demonstrate that the technique can correctly identify complexes and reliably evaluate equilibrium constants for hetero-association.


StCT2, a new antibacterial peptide characterized from the venom of the scorpion Scorpiops tibetanus.
Bacterial infection poses an increasing threat to global public health and new types of antibacterial agents are urgently needed to respond to the threat. Scorpion venom contains series of bioactive peptides, among which antibacterial peptide is an important part. Herein, a new antimicrobial peptide StCT2 was characterized from the venomous gland cDNA library of the Scorpiops tibetanus. The full-length cDNA of StCT2 is 369 nucleotides encoding the precursor that contains a putative 24 residues signal peptide, a presumed 14 residues mature peptide, and a putative 37 residues acidic propeptide at the C-terminus. The minimal inhibition concentrations (MICs) of StCT2 for Staphylococcus aureus were 6.25-25μg/ml, including antibiotic-resistant strains such as methicillin resistant S. aureus (MRSA). StCT2 was further found to show high in vivo antimicrobial activity by an S. aureus infection mouse model. StCT2 exerted its antimicrobial activity via a rapid bactericidal mechanism. Taken together, these results demonstrate the efficacy and general mechanism of StCT2 antimicrobial action and the therapeutic potential of StCT2 as a new antimicrobial peptide.


The gut microbiome: the origin of disease?
The gut microbiome in humans is considered a 'virtual organ'. They play an important part in pathophysiological and physiological processes, and regulate host homeostasis. Changes to their delicate composition can have a damaging effect on their hosts. In this article, Sarah Jane Palmer details how these microorganisms impact the human body and how they can be well-regulated.


Testosterone signaling in T cells and macrophages.
This review summarizes data about non-genomic actions of testosterone on murine malaria, T cells and macrophages produced by our group during the last 15 years. In C57BL/10 mice, testosterone induces a lethal outcome of blood stage infections with Plasmodium chabaudi which normally takes a self-healing course controlled by genes of the H-2 complex and the non-H-2 background. This suppressive effect of testosterone is mediated neither via the classic intracellular androgen receptor (AR) response nor, after conversion of testosterone to estradiol, via the estrogen receptor. Testosterone acts non-genomically, i.e. through surface receptors, on murine T cells and macrophages, which becomes evident as a rapid rise in the intracellular free Ca(2+) concentration ([Ca(2+)](i)). In T cells, this rise reflects predominantly influx of extracellular Ca(2+), while it is predominantly due to release of Ca(2+) from intracellular Ca(2+)-stores in macrophages. The testosterone-induced rise in [Ca(2+)](i) of both macrophages and T cells is not inhibited by the AR-blocker cyproterone, and it is also inducible by the plasma membrane impermeable ligand testosterone-BSA. The surface receptors initiate a transcription-independent signaling pathway of testosterone. Currently, we are trying to isolate testosterone surface receptors and to investigate a possible cross-talk of non-genomic testosterone signaling with other genotropic signaling pathways.


Forgetting in H.M.: a second look.
Forgetting was assessed in the amnesic patient H.M. using forced-choice and yes-no picture recognition at four delay intervals: 10 min, 24 hr, 72 hr, and 1 week after learning. In order to make H.M.'s initial recognition performance comparable to that of control subjects who viewed each slide for 1 sec, H.M. viewed each slide for 20 sec. H.M. displayed normal forgetting in forced-choice and yes-no recognition, although he was impaired in yes-no recognition at the 24-hr delay interval. These data contradict Huppert and Piercy's hypothesis that medical temporal-lobe pathology is associated with rapid forgetting.


Plasma tRNA Fragments Derived from 5' Ends as Novel Diagnostic Biomarkers for Early-Stage Breast Cancer.
Small RNAs derived from tRNAs are attracting considerable attention; however, the effects of tRNA-derived fragments (tRFs) and tRNA halves (tiRNAs) as biomarkers have not been investigated in early-stage breast cancer (EBC). The study aimed to explore whether tRFs and tiRNAs could be detected in plasma and whether they could serve as diagnostic biomarkers. The study was conducted in four phases. Thirty tRFs and tiRNAs were selected by high-throughput sequencing in screening phase and then assessed in training, testing, and external validation phases by qRT-PCR. Six tRFs (tRF-Glu-CTC-003, tRF-Gly-CCC-007, tRF-Gly-CCC-008, tRF-Leu-CAA-003, tRF-Ser-TGA-001, and tRF-Ser-TGA-002) were found significantly downregulated in plasma samples of patients with EBC compared with normal controls, and all were derived from 5' ends of tRNAs. Patients with HER2+ EBC with low expression levels of tRF-Glu-CTC-003 were related to worse disease-free survival and overall survival. The identified tRFs were further examined in cell supernatants, exosomes isolated from plasma, and tissues. In conclusion, our study identified six tRFs from the 5' ends of tRNAs as novel diagnostic biomarkers for EBC, providing additional evidence for, and a better understanding of, circulating tRFs and EBC.


Prevalence of joint-specific osteoarthritis and joint pain in British Columbia, Canada.
The objective is to determine the prevalence of self-reported physician-diagnosed osteoarthritis (OA) and musculoskeletal symptoms (pain, stiffness or discomfort) in specific joints among adults in British Columbia (BC), Canada. We carried out a cross-sectional mixed-mode survey in a random population sample of persons 18 years of age and older. Estimates were weighted to reflect the age and sex distribution of the population of BC. We obtained responses from 2,233 individuals. Overall, 18.4% (95% CI 16.8-20.1) of the adult population reported OA. Of those, more than 40% had OA in multiple sites. Prevalence ranged from 8.8% (95% CI 7.6-10.1) in the knee to 2.7% (2.1-3.5) in the foot. One-year prevalence of symptoms ranged from 49.1% (47.0-51.2) in the lower back to 23.3% (21.5-25.1) in the hip. Females reported more symptoms and OA than males in all joints. The most common site of self-reported physician-diagnosed OA in BC is the knee, but OA in the hands, hips, and feet is also common. Having OA in one joint is a strong predictor of OA in other joints.


Spatial and temporal effects on severe acute respiratory coronavirus virus 2 (SARS-CoV-2) contamination of the healthcare environment.
The spatial and temporal extent of severe acute respiratory coronavirus virus 2 (SARS-CoV-2) environmental contamination has not been precisely defined. We sought to elucidate contamination of different surface types and how contamination changes over time. We sampled surfaces longitudinally within COVID-19 patient rooms, performed quantitative RT-PCR for the detection of SARS-CoV-2 RNA, and modeled distance, time, and severity of illness on the probability of detecting SARS-CoV-2 using a mixed-effects binomial model. The probability of detecting SARS-CoV-2 RNA in a patient room did not vary with distance. However, we found that surface type predicted probability of detection, with floors and high-touch surfaces having the highest probability of detection: floors (odds ratio [OR], 67.8; 95% credible interval [CrI], 36.3-131) and high-touch elevated surfaces (OR, 7.39; 95% CrI, 4.31-13.1). Increased surface contamination was observed in room where patients required high-flow oxygen, positive airway pressure, or mechanical ventilation (OR, 1.6; 95% CrI, 1.03-2.53). The probability of elevated surface contamination decayed with prolonged hospitalization, but the probability of floor detection increased with the duration of the local pandemic wave. Distance from a patient's bed did not predict SARS-CoV-2 RNA deposition in patient rooms, but surface type, severity of illness, and time from local pandemic wave predicted surface deposition.


Cuprizone-Induced Demyelination in Mouse Hippocampus Is Alleviated by Ketogenic Diet.
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). Recently, ketogenic diet (KD) supplementation has attracted great interest. Therefore, we established the cuprizone (CPZ)-induced demyelination mouse model to investigate the possible neuroprotective effect of KD on the hippocampus of mice. We found that KD significantly elevated the level of serum β-hydroxybutyric acid, improved behavioral and motor abnormalities, and impaired the spatial learning and memory of CPZ-induced demyelination mice. Meanwhile, KD lessened the hippocampal demyelination by enhancing the expression of mature oligodendrocytes (OLs), which was revealed by the elevated expression of MBP and CNPase, as well as the luxol fast blue-staining intensity. Furthermore, KD inhibits the activation of microglia (especially M1-like microglia) and reactive astrocytes. Interestingly, KD attenuated the CPZ-induced oxidative stress by decreasing the malondialdehyde (MDA) content and restoring the glutathione (GSH) levels. In addition, the double immunofluorescence staining revealed that KD enhanced the expression of SIRT1 in astrocytes, microglia, and mature oligodendrocytes. Concomitantly, Western blot demonstrated that KD increased the expression of SIRT1, phosphorylated-AKT, mTOR, and PPAR-γ. In conclusion, KD exerted a neuroprotective effect on CPZ-induced demyelination mice, and this activity was associated with the modulation of the SIRT1/PPAR-γ and SIRT1/P-Akt/mTOR pathways.


Optimization of laboratory cultivation conditions for the synthesis of antifungal metabolites by bacillus subtilis strains.
In order to achieve the optimal number of colony forming units and a high level of antifungal metabolites synthesis, we carried out the periodic cultivation of the Bacillus subtilis BZR 336 g and Bacillus subtilis BZR 517 strains at various pH and temperature levels. In the experiment for determining the optimal temperature, the maximum titer of B. subtilis BZR 336 g bacterium (1.6-1.7 × 109 CFU/ml) was recorded at a cultivation temperature of 20-25 °C. For B. subtilis BZR 517 strain, the temperature turned out to be optimal at 30 °C: the titer was 8.9 × 108 CFU/ml. The maximum antifungal activity of B. subtilis BZR 336 g strain against the test culture of Fusarium oxysporum var. orthoceras was observed at a cultivation temperature of 20-25 °C; for B. subtilis BZR 517 strain, 25-30 °C. When determining the optimal pH level, it was found that a high titer of B. subtilis BZR 336 g strain cells was determined at pH 8.0 (2.7 × 109 CFU/ml), for B. subtilis BZR 517 strain it was at pH 6.0-8.0 (1.0 × 109 CFU/ml). The maximum antifungal activity was noted with the same indicators. Chromatographic and bioautographic analyses suggest that the synthesized antifungal metabolites belong to surfactin and iturin A. The data obtained in this research can be used in the development of the technology for the production of effective biofungicides to protect crops against Fusarium pathogens.


Uncertainty and expectancy deviations require cortico-subcortical cooperation.
In a dynamic and uncertain environment it is beneficial to learn the causal structure of the environment in order to minimize uncertainty. This requires determining estimates of probable outcomes, which will guide expectations about incoming information. One key factor in this learning process is to detect whether an unexpected event constitutes a low probability, but valid outcome, or an outright error. The present 7T-fMRI study investigated the role of subcortical structures in regulating this probabilistic inferential learning process. A new task was designed, in which participants learned to calculate the value, and therefore to anticipate the outcome of different visual sequences. Three types of sequences provided unambiguous, ambiguous, and incongruent contextual evidence and each sequence had two outcomes, which differed in their probability of occurrence. We hypothesized that subcortical regions are necessary when expectations are violated, and that their involvement will depend on the nature of the unexpected event. The results show increased dorsomedial striatal and thalamic activation for less probable sequences; in addition, ambiguous sequences also display larger activation in the red nuclei. Incongruent sequences displayed a pattern of subcortical activation restricted to the dorsolateral and the posterior dorsomedial striatum. These results confirm that different subcortical structures regulate uncertainty and expectancy deviations; this is crucial not only for learning to predict events in the environment, but also for flexible cognitive control in general.


White matter alteration of the cingulum in Parkinson disease with and without dementia: evaluation by diffusion tensor tract-specific analysis.
In PD, the neurodegenerative process begins in the brain stem and extends to the limbic system and finally into the cerebral cortex. We used diffusion tensor tractography to investigate the FA of the cingulate fiber tracts in patients with PD with and without dementia. Fifteen patients with PD, 15 patients with PDD, and 15 age-matched healthy controls underwent diffusion tensor imaging with a 3T MR imager. Diffusion tensor tractography images of the anterior and posterior cingulate fiber tracts were generated. Mean diffusivity and FA were measured along the tractography of the anterior and posterior cingulate fiber tracts. One-way ANOVA with the Scheffé post hoc test was used to compare results among the groups. FA was significantly lower in patients with PDD than in healthy controls in both the anterior and the posterior cingulate fiber tracts (P = .003, P = .015) and significantly lower in patients with PD than in healthy controls (P = .003) in the anterior cingulate fiber tract. There were no significant mean diffusivity differences among the groups. MMSE and FA values of the anterior cingulate fiber tracts in patients with PDD were significantly correlated (r = 0.633, P < .05). The reduced FA in patients with PD and PDD might reflect neuropathologic changes such as Lewy body pathology in the cingulate fibers. This abnormality might contribute to the dementing process in PD.


Dietary accumulation of hexabromocyclododecane diastereoisomers in juvenile rainbow trout (Oncorhynchus mykiss) I: bioaccumulation parameters and evidence of bioisomerization.
Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to three diastereoisomers (alpha, beta, gamma) of hexabromocyclododecane (C12H18Br6) via their diet for 56 d followed by 112 d of untreated food to examine bioaccumulation parameters and test the hypothesis of in vivo bioisomerization. Four groups of 70 fish were used in the study. Three groups were exposed to food fortified with known concentrations of an individual diastereoisomer, while a fourth group were fed unfortified food. Bioaccumulation of the gamma-diastereoisomer was linear during the uptake phase, while the alpha- and beta-diastereoisomers were found to increase exponentially with respective doubling times of 8.2 and 17.1 d. Both the beta- and the gamma-diastereoisomers followed a first-order depuration kinetics with calculated half-lives of 157 +/- 71 and 144 +/- 60 d (+/-1 x standard error), respectively. The biomagnification factor (BMF) for the alpha-diastereoisomer (BMF = 9.2) was two times greater than the beta-diastereoisomer (BMF = 4.3); the large BMF for the beta-diastereoisomer is consistent with this diastereoisomer dominating higher-trophic-level organisms. Although the BMF of the beta-diastereoisomer suggests that it will biomagnify, it is rarely detected in environmental samples because it is present in small quantities in commercial mixtures. Results from these studies also provide evidence of bioisomerization of the beta- and gamma-diastereoisomers. Most importantly, the alpha-diastereoisomer that was recalcitrant to bioisomerization by juvenile rainbow trout in this study and known to be the dominant diastereosiomer in fish was bioformed from both the beta- and the gamma-diastereoisomers. To our knowledge, this is the first report of bioisomerization of a halogenated organic pollutant in biota.


Multi-task learning for Chinese clinical named entity recognition with external knowledge.
Named entity recognition (NER) on Chinese electronic medical/healthcare records has attracted significantly attentions as it can be applied to building applications to understand these records. Most previous methods have been purely data-driven, requiring high-quality and large-scale labeled medical data. However, labeled data is expensive to obtain, and these data-driven methods are difficult to handle rare and unseen entities. To tackle these problems, this study presents a novel multi-task deep neural network model for Chinese NER in the medical domain. We incorporate dictionary features into neural networks, and a general secondary named entity segmentation is used as auxiliary task to improve the performance of the primary task of named entity recognition. In order to evaluate the proposed method, we compare it with other currently popular methods, on three benchmark datasets. Two of the datasets are publicly available, and the other one is constructed by us. Experimental results show that the proposed model achieves 91.07% average f-measure on the two public datasets and 87.05% f-measure on private dataset. The comparison results of different models demonstrated the effectiveness of our model. The proposed model outperformed traditional statistical models.


Mood effects of alternate-day corticosteroid therapy in patients with systemic lupus erythematosus.
Mood and cognition were assessed on consecutive days in 18 female patients with systemic lupus erythematosus on alternate-day corticosteroid therapy. No overall differences in mood and cognition were observed between the on- and off-medication days. However, 10 patients showed marked worsening or improvement of either depression or anxiety on their off-medication day. Two weeks of prospective behavioral ratings confirmed the observed mood changes in several patients. The clinical and theoretical implications of these findings are discussed.


MR imaging of muscular sarcoidosis after steroid therapy.
The nodular type of muscular sarcoidosis has been known to show characteristic MR findings: however, MR imaging features after steroid therapy have not been reported. A 48-year-old man with nodular type of muscular sarcoidosis is reported. Prior to steroid therapy, axial MR images showed peripheral increased signal intensity and central star-shaped decreased signal intensity. Coronal images showed an inner stripe of decreased signal intensity and outer stripes of increased signal intensity. After steroid therapy, axial images showed only the central star-shaped area of decreased signal intensity. Coronal images showed only the inner stripe of decreased signal intensity. It is important to know that the central area will continue to exist after steroid therapy.


Pregnancy associated hormones modulate the cytokine production but not the phenotype of PBMC-derived human dendritic cells.
Dendritic cells (DC) play a central role in initiating and polarizing immune responses. As effects of pregnancy associated hormones on phenotype and function of DC are unknown, our objective was to test the influence of progesterone, beta-estradiol and betaHCG on immature (iDC) and mature (mDC) DC. DC generated from peripheral-blood-monocytes were exposed to different doses of hormones. DC phenotype was determined by FACS-analysis of surface marker expression (CD40, CD86, CD83 and HLA-DR). Modifications in the secretion of cytokines (IL12p70, IL-18, IL-10, IL-6, TNFalpha) and chemokines (MDC, IL-8) were analysed by ELISA. T cell stimulatory capacity of mDC was assessed by mixed lymphocyte reaction. Incubation with progesterone or estradiol resulted in a significant upregulation of IL-10 production by iDC and mDC. Combinations of progesterone and betaHCG or estradiol respectively induced a significant decrease in production of IL-18 by mDC. No significant changes could be observed in surface marker expression or T cell stimulatory capacity, neither in cultures of DC matured under influence of progesterone, estradiol nor betaHCG. PBMC-derived DC seem to be relatively stable against the influence of pregnancy associated hormones apart from particular effects on cytokine production which partly could contribute to the modification of immune responses observed in normal early pregnancy.