A hydrogel-based versatile screening platform for specific biomolecular recognition in a well plate format.
Precise determination of biomolecular interactions in high throughput crucially depends on a surface coating technique that allows immobilization of a variety of interaction partners in a non-interacting environment. We present a one-step hydrogel coating system based on isocyanate functional six-arm poly(ethylene oxide)-based star polymers for commercially available 96-well microtiter plates that combines a straightforward and robust coating application with versatile bio-functionalization. This system generates resistance to unspecific protein adsorption and cell adhesion, as demonstrated with fluorescently labeled bovine serum albumin and primary human dermal fibroblasts (HDF), and high specificity for the assessment of biomolecular recognition processes when ligands are immobilized on this surface. One particular advantage is the wide range of biomolecules that can be immobilized and convert the per se inert coating into a specifically interacting surface. We here demonstrate the immobilization and quantification of a broad range of biochemically important ligands, such as peptide sequences GRGDS and GRGDSK-biotin, the broadly applicable coupler molecule biocytin, the protein fibronectin, and the carbohydrates N-acetylglucosamine and N-acetyllactosamine. A simplified protocol for an enzyme-linked immunosorbent assay was established for the detection and quantification of ligands on the coating surface. Cell adhesion on the peptide and protein-modified surfaces was assessed using HDF. All coatings were applied using a one-step preparation technique, including bioactivation, which makes the system suitable for high-throughput screening in a format that is compatible with the most routinely used testing systems.


Protein-protein interactions involving congenital cataract T5P gammaC-crystallin mutant: a confocal fluorescence microscopy study.
The human lens crystallin gene CRYGC T5P is associated with Coppock-like cataract and has a phenotype of a dust-like opacity of the fetal lens nucleus and deep cortical region. Previous in vitro mutation studies indicate that the protein has changed conformation, solubility, and stability, which may make it susceptible to aggregation, as seen in cataractous lens and cell culture expression. To investigate the mechanisms leading to these events, we studied protein-protein interactions using confocal fluorescence resonance energy transfer (FRET) microscopy. The method detects protein-protein interactions in the natural environment of living cells. Crystallin genes (CRYGC T5P, CRYGC, and CRYAA) were fused to either the green fluorescence protein (GFP) or red fluorescence protein (DsRED or RFP) vector. Each of the following GFP-RFP (donor-acceptor) plasmid pairs was cotransfected into HeLa cells: gammaC-gammaC, gammaC-gammaCT5P, gammaCT5P-gammaCT5P, alphaA-gammaC, and alphaA-gammaCT5P. After culture, confocal fluorescence cell images were taken. Protein-protein interactions in the form of net FRET were evaluated. The confocal fluorescence images show that cells expressing T5P gammaC-crystallin contain many protein aggregates, but cells co-expressing with either gammaC- or alphaA-crystallin reduce the aggregation considerably. FRET determination indicates that gammaCT5P-gammaCT5P shows less protein-protein interaction than either gammaC-gammaC or gammaC-gammaCT5P. Cotransfection with alphaA-crystallin (alphaA-gammaC or alphaA-T5PgammaC) increases nFRET compared with gammaC-gammaC or gammaC-T5PgammaC. Our results demonstrate that T5P gammaC-crystallin shows more protein aggregates and less protein-protein interaction than WT gammaC-crystallin. Chaperone alphaA-crystallin can rescue T5P gammaC-crystallin from aggregation through increased protein interaction. The formation of congenital cataract may be due to reduced protein-protein interactions and increased aggregation from an insufficient amount of alpha-crystallin for protection.


Phase I study on the pharmacokinetics and tolerance of ZT-1, a prodrug of huperzine A, for the treatment of Alzheimer's disease.
Huperzine A isolated from the Chinese herb Huperzia serrata (Thunb) Trev is a novel reversible and selective AChE inhibitor. The aim of this study was to evaluate the pharmacokinetics and tolerance of single and multiple doses of ZT-1, a novel analogue of huperzine A, in healthy Chinese subjects. This was a double-blinded, placebo-controlled, randomized, single- and multiple-dose study. For the single-dose study, 9 subjects were randomly divided into 3 groups receiving ZT-1 (0.5, 0.75 or 1 mg, po) according to a Three-way Latin Square Design. For the multiple-dose study, 9 subjects receiving ZT-1 (0.75 mg/d, po) for 8 consecutive days. In the tolerance study, 40 subjects were randomly divided into 5 groups receiving a single dose of ZT-1 (0.5, 0.75, 1, 1.25 or 1.5 mg, po). Plasma and urine concentrations of ZT-1 and Hup A were determined using LC-MS/MS. Pharmacokinetic parameters, including Cmax, AUC0-72 h and AUC0-∞ were calculated. Tolerance assessments were conducted throughout the study. ZT-1 was rapidly absorbed and converted into huperzine A, thus the plasma and urine concentrations of ZT-1 were below the limit of quantification (<0.05 ng/mL). After single-dose administration of ZT-1, the mean tmax of huperzine A was 0.76-0.82 h; the AUC0-72 h and Cmax of huperzine A showed approximately dose-proportional increase over the dose range of 0.5-1 mg. After the multiple-dose administration of ZT-1, a steady-state level of huperzine A was achieved within 2 d. No serious adverse events were observed. ZT-1 is a pro-drug that is rapidly absorbed and converted into huperzine A, and ZT-1 is well tolerated in healthy Chinese volunteers.


The effect of temporal lobe surgery on electrodermal activity: implications for an organic hypothesis in the aetiology of schizophrenia.
It has recently been suggested that the bilateral asymmetry of electrodermal activity (EDA) reported in schizophrenia may be related to unilateral temporal lobe dysfunction. To test this hypothesis, 3 aspects of EDA--skin conductance level (SCL), number of spontaneous fluctuations (SF), and skin conductance response (SCR)--were measured bilaterally in 10 patients who had undergone unilateral temporal lobectomy. No differences could be detected between the operated and non-operated sides within the patient group, nor between the patient and control groups. The implications of these findings are discussed.


The DNA co-vaccination using Sm antigen and IL-10 as prophylactic experimental therapy ameliorates nephritis in a model of lupus induced by pristane.
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies such as anti-Sm. Studies in patients with SLE and murine models of lupus reveal that the most critical anti-Sm autoantibodies are predominantly direct against D1(83-119), D2, and B´/B epitopes. The present study aimed to analyze the induction of antigen-specific tolerance after prophylactic immunization with a DNA vaccine encoding the epitopes: D183-119, D2, B´/B, and B´/BCOOH in co-vaccination with IFN-γ or IL-10 in a murine model of lupus induced by pristane. To obtain endotoxin-free DNA vaccines, direct cloning techniques using pcDNA were performed: D183-119, D2, B´/B, B´/BCOOH, IFN-γ, or IL-10. Lupus was induced by 0.5 mL of pristane via intraperitoneal in BALB/c female mice. Immunoprecipitation with K562 cells was metabolically labeled with 35S and ELISA to detect serum antibodies or mice IgG1, IgG2a isotypes. ELISA determined IL-10 and IFN-γ from splenocytes supernatants. Proteinuria was assessed monthly, and lupus nephritis was evaluated by immunofluorescence, and electron microscopy. The prophylactic co-vaccination with D2/IL-10 reduced the expression of kidney damage observed by electron microscopy, direct immunofluorescence, and H & E, along with reduced level of anti-nRNP/Sm antibodies (P = 0.048). The prophylactic co-vaccination of IL-10 with D2 in pristane-induced lupus ameliorates the renal damage maybe by acting as prophylactic DNA tolerizing therapy.


Identification of high priority focal activations in persistent atrial fibrillation using a novel mapping strategy.
Focal activation is believed to be an atrial fibrillation (AF) driver; however, little is known about whether all focal activations are necessary for AF persistence. The purpose of this study was to assess the electrical nature of focal activation and identify high-priority focal activations using a novel mapping system (CARTOFINDER). Thirty-five patients with persistent AF who underwent catheter ablation were assessed. Cycle length (CL) and CL standard deviation (CLSD) on unipolar recordings and voltage amplitude and electrogram morphologies on bipolar recordings were evaluated at all points of interest. The most frequent CL at each mapping site was defined as the dominant CL. We identified dominant focal activations (DFAs) that had a shorter dominant CL on the integrated CARTOFINDER map. The effect of elimination of DFAs on AF maintenance was assessed by the composite endpoint (termination to sinus rhythm, organization of the rhythm to atrial tachycardia, and AF CL slowing). In all, 450 focal activations were identified among 10,868 points, and 50.4% of focal activations were DFAs. Focal activations showed relatively long CL and regularity with short CLSD. Most focal activations showed an isoelectric baseline and were located outside of the fractionated electrogram area. Both DFAs and non-DFAs were typically observed in the normal voltage range. Elimination of DFAs was achieved in 19 (54.3%) patients, with a remarkable impact on AF maintenance (68.4% vs. 25.0%, p = 0.018). In conclusion, DFAs may play an important role in AF maintenance and could be an attractive therapeutic target for AF.


Flow cytometric measurement of apoptosis and necrosis in cryopreserved PBPC concentrates from patients with malignant diseases.
The number of viable precursor cells actually reinfused into patients after high-dose chemotherapy is one of the most clinically important variables determining graft success or failure. A modified, previously described flow cytometric method based on annexin V staining was therefore applied to assess the degree of apoptosis and necrosis in cryopreserved PBPC concentrates from patients with malignant diseases. Twenty-two samples of unmanipulated cryopreserved PBPC concentrates were analyzed by flow cytometry. The samples were triple-stained with anti-CD34 PE, annexin V-FITC and actinomycin D, which enabled the separation of viable, early apoptotic and late apoptotic/necrotic CD34(+) precursor cells. Apotosis and necrosis were also measured in the total cell population of the concentrates. Eighty-one percent (range 49-97) of the CD34(+) cells were viable, while 7% (range 1-15) were early apoptotic and 12% (range 2-36) were late apoptotic/necrotic after freeze/thaw. There was no difference in apoptosis and necrosis in CD34(+) cells harvested from mildly pretreated patients with multiple myeloma and heavily pre-treated patients with non-Hodgkin's lymphoma. Apoptosis and necrosis were higher in the total mature cell population of the concentrates. Thirty-two percent (range 7-69) of the cells were apoptotic and 33% (range 12-60) were necrotic. We conclude that flow cytometric analysis of annexinV/actinomycin D binding in PBPC concentrates is a simple technique that can give additional information of the viability status of the cells post thaw. The present study confirms the relative robustness of human CD34(+) precursor cells concerning the freeze/thaw procedure, which are carried out in daily clinical practice.


Large-scale climate patterns offer preseasonal hints on the co-occurrence of heat wave and O3 pollution in China.
Heat waves and air pollution extremes exert compounding effects on human health and food security and may worsen under future climate change. On the basis of reconstructed daily O3 levels in China and meteorological reanalysis, we found that the interannual variability of the frequency of summertime co-occurrence of heat wave and O3 pollution in China is regulated mainly by a combination of springtime warming in the western Pacific Ocean, western Indian Ocean, and Ross Sea. These sea surface temperature anomalies impose influences on precipitation, radiation, etc., to modulate the co-occurrence, which were also confirmed with coupled chemistry-climate numerical experiments. We thus built a multivariable regression model to predict co-occurrence a season in advance, and correlation coefficient could reach 0.81 (P < 0.01) for the North China Plain. Our results provide useful information for the government to take actions in advance to mitigate damage from these synergistic costressors.


Community pharmacist-led medication review procedures across Europe: Characterization, implementation and remuneration.
Pharmaceutical Care Network Europe (PCNE) proposed a definition and classification system (type 1, 2a, 2b, 3) for medication review in 2016. However, to date, a description of the implementation and remuneration of such procedures across Europe is lacking. The aim of this study was to describe the medication review procedures and the level of implementation and remuneration in community pharmacies across Europe. An online survey was developed to characterize medication review procedures (PCNE classification), level of implementation (considering regional or national) and remuneration by a third party. This survey was sent to a purposive sample of three individuals per country, with a working background in community pharmacy, pharmacy practice research, or health policy to ensure reliable data. Data triangulation was used and consensus sought between the responses. Data were received from 34 out of 44 targeted European countries (November 2016-October 2017) [response rate = 77%]. Overall, 55.9% of the countries provided at least one type of medication review as an implemented service or project. Type 1 medication review (based on the medication history) was provided in 13 countries, type 2a (medication history + patient interview) in 14, type 2b (medication history + clinical data) in two, and type 3 medication review (medication history + patient interview + clinical data) in four countries. Ten of the mentioned services or projects were remunerated by a third-party. Substantial heterogeneity was observed across Europe in various aspects, including the procedures, implementation level and remuneration obtained. Type 1 and 2a medication review services seem to be more feasible to implement in the community pharmacy than type 2b and 3. A large number of medication review projects were ongoing in community pharmacies, which suggests that new medication review services could become implemented in the coming years.


Novel calcium-binding GTPase (AtCBG) involved in ABA-mediated salt stress signaling in Arabidopsis.
We have identified a novel Ca(2+)-signal sensing GTPase (643 amino acid residues with an estimated molecular mass of 79 kDa) from the Arabidopsis genome database. This protein contains a RHO-like GTPase domain at the N-terminus (15-184 amino acids) and two calcium-binding EF-hand motifs (199-227 and 319-347 amino acids, respectively). It has the capability to bind calcium and hydrolyze GTP; in addition, its GTPase activity is regulated by changes in Ca(2+) concentration. The expression of this gene was induced by ABA and salt stresses, and specific knock-out mutants were highly sensitive to ABA and salt treatments. These findings suggest that this protein is a novel ABA- and salt stress-related Ca(2+) signal transducer.


Controllable synthesis of nanostructured ZnCo2O4 as high-performance anode materials for lithium-ion batteries.
Nanostructured ZnCo2O4 anode materials for lithium-ion batteries (LIBs) have been successfully prepared by a two-step process, combining facile and concise electrospinning and simple post-treatment techniques. Three different structured ZnCo2O4 anodes (nanoparticles, nanotubes and nanowires) can be prepared by simply adjusting the ratio of metallic salt and PVP in the precursor solutions. Charge-discharge tests and cyclic voltammetry (CV) have been conducted to evaluate the lithium storage performances of ZnCo2O4 anodes, particularly for ZnCo2O4 nanotubes obtained from a weight ratio 2 : 4 of metallic salt and PVP polymer in the precursor solution. Remarkably, ZnCo2O4 nanotubes exhibit high specific capacity, good rate property, and long cycling stability. Reversible capacity is still maintained at 1180.8 mA h g-1 after 275 cycles at a current density of 200 mA g-1. In case of rate capability, even after cycling at the 2000 mA g-1 current density, the capacity could recover to 684 mA h g-1. The brilliant electrochemical properties of the ZnCo2O4 anodes make them promising anodes for LIBs and other energy storage applications.


Medial column procedures in the correction of adult acquired flatfoot deformity.
AAFD is a complex problem with a wide variety of treatment options. No single procedure or group of procedures can be applied to all patients with AAFD because of the variety of underlying etiology and grades of deformity. As the posture of the foot progresses into hindfoot valgus and forefoot abduction through attenuation of the medial structures of the foot, the medial column begins to change shape. The first ray elevates and the joints of the medial column may begin to collapse. Careful physical examination and review of weight-bearing radiographs determines which patients have an associated forefoot varus deformity that may require correction at the time of flatfoot reconstruction. Correction of an AAFD requires a combination of soft-tissue procedures to restore dynamic inversion power and bony procedures to correct the hindfoot and midfoot malalignments. If after these corrections forefoot varus deformity remains, the surgeon should consider use of a medial column procedure to recreate the "triangle of support" of the foot that Cotton described.5 If the elevation of the medial column is identified to be at the first NC or the first TMT joint, then the joint should be carefully examined for evidence of instability, hypermobility, or arthritic change. If none of these problems exist, then the surgeon can consider use of the joint-sparing Cotton medial cuneiform osteotomy to correct residual forefoot varus. However, if instability, hypermobility, or arthritic change is present, then the surgeon should consider use of an arthrodesis of the involved joint to correct residual forefoot varus. Either procedure provides a safe and predictable correction to the medial column as part of a comprehensive surgical correction of AAFD.


Tropisetron attenuated the anxiogenic effects of social isolation by modulating nitrergic system and mitochondrial function.
Early social isolation stress (SIS) is associated with the occurrence of anxiety behaviors. It seems interaction between the nitrergic system and mitochondrial function plays a role in mediating the anxiety-like behaviors. In this study, we aimed to investigate the anxiolytic effects of tropisetron in animal model of SIS and we try to illustrate the possible role of nitrergic system and mitochondrial function. We applied early social isolation paradigm to male NMRI mice. Animals treated with various doses of tropisetron, nitric oxide agents or their combination and anxiety-like behaviors of animals were assessed using valid behavioral tests including elevated plus maze (EPM), open-field test (OFT) and hole-board test (HBT) in their adulthood. Effects of housing conditions and drug treatments on the mitochondrial function were investigated in the hippocampus by assessing the ATP, GSH, ROS and nitrite levels. Anxiogenic effects of early SIS were assessed in the EPM, OFT, and HBT. Also, SIS disrupted mitochondrial function and caused oxidative stress in the hippocampus of stressed animals. Tropisetron showed an anxiolytic effect in the stressed mice. Also, these effects were mediated by nitrergic system by affecting mitochondrial function and modulating the oxidative stress. L-arginine, a nitric oxide precursor, abolished the anxiolytic effects of tropisetron in the behavioral tasks and blocked the protective effects of it against mitochondrial and oxidative challenge. Our results demonstrated tropisetron attenuated the anxiogenic effects of SIS by mitigation of the negative effects of nitric oxide on mitochondrial function.


Using a multimedia presentation to improve patient understanding and satisfaction with informed consent for minimally invasive vascular procedures.
As vascular procedures become more complex, patient understanding of their treatment(s) can become more difficult. We wished to evaluate the utility of multimedia presentations (MPs) to improve patient understanding of their vascular interventions. Patients undergoing endovascular aneurysm repair (EVAR), peripheral angioplasty, Hickman catheter and peripherally inserted central catheter (PICC) insertion were randomized into a control group receiving traditional verbal consent, and a MP group that were shown a two minute simplified video of their procedure on an iPad™ computer in addition to the traditional verbal consent. After obtaining consent, all patients completed a questionnaire assessing their comprehension of the procedure, and satisfaction with the consent process. Satisfaction was rated on a 5 point Likert scale with 5 being 'very helpful' in understanding the procedure. Ninety-three patients were recruited for this study, 62% of which were male. The intervention significantly increased total comprehension in all procedure types controlling for procedure type (multimedia vs. control; F = 9.14, P = .003). A second ANOVA showed there was a significant main effect by intervention (F = 44.06, p < .000) with those in the intervention group showing higher overall satisfaction scores after controlling for surgery type. This study suggests that patients find the use of MP during the consent process to be helpful in patient understanding and that there is improved satisfaction. Given the rapid rate of innovation in vascular interventions, increased regular use of MPs to help patients understand their procedures would be beneficial in the care of patients undergoing vascular interventions.


Total mercury, organic mercury, copper, manganese, selenium, and zinc in Sarda sarda from the Gulf of Genoa.
The total mercury content of the Atlantic bonito (Sarda sarda) increases with increasing size of the specimen, as shown by other authors for other marine organisms (Renzoni et al., 1973; Buffoni et al., 1982; Bernhard and Andreae, 1984; Thibaud, 1986). Organic mercury content also increases with the size of the specimen, whereas inorganic mercury remains constant, which is in agreement with the prediction made on another species of Sgombridae (Bernhard, 1985). This kind of mercury partitioning has not, to our knowledge, been reported for other species of fish. It is interesting to note that the mercury concentration in the gonads also increases with the size of the specimen. A very small difference between the total mercury content of white and dark muscle was observed, while for selenium, zinc, copper, and manganese the concentrations are lower in white muscle than in dark muscle.


Intronic variants of LGALS13 gene encoding placental protein (PP13) are linked with increased risk of infection-associated spontaneous preterm birth.
Spontaneous preterm birth (sPTB) is a global health issue. Studies suggest infection and infection-based inflammatory responses are major risk factors for sPTB. Considering the important role of anti-inflammatory proteins in pregnancy, the study aimed to find the association between anti-inflammatory LGALS13 gene variants IVS2-22 A/G (rs2233706) and IVS3+72 T/A (rs2233708) and the risk of sPTB during Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum infection in Indian population. Placental samples of 160 sPTB and 160 term women were collected. Pathogens were detected by PCR. The genotyping of LGALS13 gene variants IVS2-22 A/G (rs2233706) and IVS3+72 T/A (rs2233708) was done by qualitative real-time PCR using allelic discrimination method (VIC- and FAM-labeled). The frequency of AG or GG genotype of LGALS13 IVS2-22A/G polymorphism (rs2233706) was 75.5% in infected sPTB cases and 14.4% in uninfected sPTB cases and 7.3% in term birth controls (p < .0001), while the frequency of TA or AA genotype of LGALS13 IVS3+72T/A polymorphism (rs2233708) was 83.6% in infected sPTB cases and 18% in uninfected sPTB cases and 12.7% in term birth controls (p < .0001). The genotypic frequencies for both the variants of LGALS13 were statistically significant (p < .0001) in the infected sPTB versus uninfected sPTB and term birth controls. Study reveals strong association between the presence of immunological gene variants LGALS13 IVS2-22 A/G (rs2233706) and LGALS13 IVS3+72 T/A (rs2233708) and risk of sPTB during C. trachomatis, M. hominis and U. urealyticum infection.


Comparison of FDG-PET/CT and MR with diffusion-weighted imaging for assessing peritoneal carcinomatosis from gastrointestinal malignancy.
To assess the accuracy of FDG-PET/CT and MR with diffusion-weighted imaging (MR-DWI) for diagnosing peritoneal carcinomatosis (PC) from gastrointestinal malignancies. Thirty consecutive patients referred for staging of gastrointestinal malignancy underwent FDG-PET/CT and MR-DWI in this retrospective study. Extent of PC was characterised by dividing the peritoneal cavity into three sites in each patient: right and left supramesocolic areas and inframesocolic level (total 90 sites). Presence of PC was confirmed either by surgery (18/30) or by follow-up (12/30). PC was confirmed in 19 patients (19/30). At a total of 90 sites, 27 showed proven PC. On a patient-based analysis, sensitivity, specificity, PPV, NPV and accuracy were respectively 84%, 73%, 84%, 73% and 80% for PET/CT and 84%, 82%, 89%, 75% and 83% for MR-DWI. On a site-based analysis, overall sensitivity and specificity of PET/CT (63%, 90%) and MR-DWI (74%, 97%) were not statistically different (P = 0.27). In the supramesocolic area, MR-DWI detected more sites involved than PET/CT (7/9 vs. 4/9). The sensitivities of PET and MR were lower for subcentimetre tumour implants (42%, 50%). Interobserver agreement was very good for PET/CT and good for MR-DWI. FDG-PET/CT and MR-DWI showed similar high accuracy in diagnosing PC. Both techniques underestimated the real extent of PC because of decreased sensitivity for subcentimetre lesions. FDG-PET/CT and MR-DWI showed similar high accuracy for diagnosing peritoneal carcinomatosis. • In the supramesocolic area, MR-DWI could be more sensitive than PET/CT. • Both techniques showed lower sensitivity for subcentimetre lesions. • Interobserver agreement was very good for PET/CT and good for MR-DWI.


Association between gross motor function and postural control in sitting in children with Cerebral Palsy: a correlational study in Spain.
Cerebral palsy (CP) is one of the causes of physical disability in children. Sitting abilities can be described using the Level of Sitting Scale (LSS) and the Gross Motor Function Classification System (GMFCS). There is growing interest in the sitting posture of children with CP owing to a stable sitting position allows for the development of eye-hand coordination, functions of the upper extremities and functional skills. Besides, in recent years researchers have tried to develop a new terminology to classify the CP as performed by the Surveillance of Cerebral Palsy in Europe (SCPE), in order to improve the monitoring of the frequency of the PC, providing a framework for research and service planning. The aim of this study was to analyse the relationship between GMFCS and LSS. The second purpose was to describe how the SCPE relates to sitting abilities with the GMFCS and LSS. The study involved 139 children with CP (range 3-18 years) from 24 educational centres. Age, gender, CP classification according to SCPE, GMFCS and LSS levels were recorded by an experienced physiotherapist. A significant inverse relationship between GMFCS and LSS score levels was found (rs = -0.86, p = 0.00). 45.3% of the children capable of leaning in any direction and of re-erecting the trunk (level VIII on the LSS) could walk without limitation (level I on the GMFCS). There were differences in the distribution of the GMFCS (χ(2)(4):50.78) and LSS (χ(2)(7): 37.15) levels and CP according to the distribution of the spasticity (p < 0.01). There was a negative correlation between both scales and a relation between sitting ability and the capacity to walk with or without technical devices. GMFCS and the LSS are useful tools for describing the functional abilities and limitations of children with CP, specially sitting and mobility. Classification based on the distribution of spasticity and the gross motor function provides clinical information on the prognosis and development of children with CP.


Measurement of left atrial appendage size by transesophageal echocardiography.
Transesophageal echocardiography (TEE) is the most common imaging method for evaluating left atrial morphology. Recent advances in 64-slice multidetector computed tomography (64-MDCT) allow accurate measurement of left atrial appendage (LAA) volume. The aim of this study was to evaluate the accuracy of LAA sizing by TEE in comparison with 64-MDCT in patients with atrial fibrillation. Electrocardiogram-gated 64-MDCT and TEE were performed within 2 days in 18 consecutive patients (63 ± 9 years old, 12 males, 5 paroxysmal atrial fibrillation) with nonvalvular atrial fibrillation. LAA area and LAA volume were measured at end-systole by TEE and 64-MDCT, respectively. The largest LAA area was measured on TEE image. Five patients were in sinus rhythm during examinations. In all patients, LAA was clearly visualized; the largest area of LAA was 9.3 ± 3.9 mm(2) and the LAA volume was 21.6 ± 7.5 ml. A significant correlation between LAA area and LAA volume was observed (p = 0.0003, r = 0.75). TEE allows a detailed evaluation of the LAA structure by two-dimensional imaging. LAA size could be evaluated by TEE despite its morphological complexity, i.e., sac-like or multilobed structure.


Tolerance and efficacy of parenterally administered penicillin-streptomycin and orally administered amoxicillin or penicillin V for prophylaxis of experimentally induced streptococcal endocarditis.
A regimen of a single intramuscular dose of penicillin G-streptomycin was compared with regimens of three oral doses of amoxicillin and two oral doses of penicillin V to prevent Streptococcus sanguis endocarditis in rabbits with experimentally induced valvular heart lesions. Challenge doses of 10(4), 10(6), and 10(8) CFU of a strain of S. sanguis highly tolerant to penicillin and amoxicillin were used. The combination of penicillin and streptomycin was the only regimen tested that provided full protection even against the highest inoculum concentration. A single oral dose of penicillin V (36 mg/kg) or amoxicillin (50 mg/kg), two oral doses of penicillin V (36 and 18 mg/kg with a 7-h interval between doses), or six oral doses of amoxicillin (50 mg/kg followed by 8.5 mg/kg at 8-h intervals) protected recipients of the lowest inoculum concentration; protection diminished with increasing inocula. In contrast, administration of two high oral doses of amoxicillin (50 mg/kg) with a 10-h interval between doses provided full protection against challenge doses of 10(4) and 10(6) CFU, preventing endocarditis in 10 (66%) of 15 recipients of 10(8) CFU. All regimens evaluated were highly effective in preventing endocarditis when rabbits were challenged with 10(4) CFU. The combination of penicillin and streptomycin was the best regimen tested. Administration of two high oral doses of amoxicillin (50 mg/kg) with a 10-h interval between doses led to significantly fewer infections when compared with the other oral regimens when rabbits were challenged with 10(6) and 10(8) CFU.


Associations between diurnal changes in Goldmann tonometry, corneal geometry, and ocular response analyzer parameters.
To investigate the possible covariations between diurnal changes in Goldmann applanation tonometry (GAT) estimates of intraocular pressure (IOP), central corneal thickness (CCT), central corneal radius, and ocular response analyzer measures of corneal hysteresis and corneal resistance factor (CRF). Twenty-five healthy normal subjects (age, 21 ± 2 years) had central corneal radius, ocular response analyzer, GAT, and CCT measurements performed in 1 eye at multiple times for more than 24 hours. Measurements were taken at 17:00 and repeated every 2 hours until 23:00. Subjects then slept uninterrupted overnight for approximately 8 hours. On awakening, readings were taken at a 20-minute frequency from 7:00 to 9:00 inclusive, followed by 2 hourly intervals until 17:00. The maximum variation in GAT for more than 24 hours was 2.5 mm Hg, and these fluctuations were predicted by the diurnal variations of CCT (0.37 mm Hg ΔGAT/10 μm CCT, P < 0.01) and CRF (0.68 mm Hg ΔGAT/mm Hg CRF, P < 0.01). The diurnal changes in corneal hysteresis and CRF both covaried with those of CCT (0.16 and 0.30 mm Hg/10 μm ΔCCT, respectively, both Ps < 0.01). None of these associations was statistically different during and after the nominal period of resolution from overnight corneal swelling (2-way interactions, all Ps > 0.05). If the IOP is assumed to be independent of corneal parameters, the clinically significant 24-hour variation in GAT being partially predicted by the changes in CCT and CRF suggests that the diurnal variation in corneal parameters affects tonometry measurements. CRF may also be dependent on the level of IOP.


A self-psychological study of a shared gambling fantasy in Eugene O'Neill's Hughie.
This paper presents an applied psychoanalytic study of Eugene O'Neill's two-character play, Hughie. Applying the constructs of self psychology, the play illustrates both the narcissistic features and the emotional and behavioral characteristics of compulsive gamblers. The study focuses particular attention on the role of narcissistic fantasies--with both grandiose and megalomaniacal features--in affecting, temporarily, the mood of the characters. Moreover, it is shown that a shared gambling fantasy--a "winner among winners"--enables them to experience a sense of camaraderie, humanness, and the illusion of kinship.


Proliferation of intestinal crypt cells by gastrin-induced ornithine decarboxylase.
To determine whether the gastrin stimulated intestinal crypt cell (IEC-6) proliferation by induction of ornithine decarboxylase (ODC). IEC-6 cells were grown in DMEM containing 50 mL x L(-1) dialyzed fetal bovine serum for 24h and then were treated with gastrin. The proliferative capability of the cells was monitored subsequently on d 1, 2, 3, and 4 after treatment with MTT assay at aborbance 570 nm.The cellular ODC mRNA expression, ODC activity, and putrescine content were examined by RT-PCR method, radiometric technique and high-performance liquid chromatography(HPLC) analysis respectively after 12h of treatment. On d1 after exposure of IEC-6 cells to pentagastrin, the proliferation increased initially and reached a peak on d3 at 250 microg x L(-1) concentration. Pentagastrin 500microg x L(-1) increased cell proliferation on day 1 and day 2, and then decreased. Compared with control group, pentagastrin 250 microg x L(-1) increased ODC mRNA level by 1.09-fold (P<0.05), ODC activity by 1.71-fold(P<0.01), and putrescine content 5.30-fold (P<0.01), respectively. Similarly, pentagastrin of 500 microg x L(-1) also increased ODC mRNA level by 1.16-fold (P<0.05), ODC activity 1.63-fold(P<0.05), and putrescine content 4.41-fold (P<0.01), respectively. But there was not significant difference between them. Gastrin is an agent which promotes IEC-6 cell proliferation involved in regulating ODC activity mechanism.


High voltage electron microscopic studies of endothelial cell tubular structures in the mouse blood-brain barrier following brain trauma.
High-voltage electron microscopy was applied to the study of endothelial cell (EC) transport of macromolecules in a murine model of blood-brain barrier injury to study the role of the EC canalicular system following brain insult. Semithick sections from mouse brains subjected to acute (2-3 h) mechanical trauma demonstrated permeation of intravenously injected horseradish peroxidase via tubular structures either (a) in the absence of lysosome-associated structures in close proximity, or (b) in association with lysosomes, dense bodies or multivesicular bodies. Our data suggest a dual-purposed system of tubules, one portion that supplies the metabolic requirements of the cell and another portion, suggested to be more limited, that opens up as a result of brain injury.


Breast-feeding and health professionals: a study in hospitals in Indonesia.
This paper presents findings on knowledge, attitudes, and practices regarding breast-feeding management in the modern health sector in Indonesia. The methodology applied was a survey which was carried out in teaching hospitals in major cities throughout Indonesia. The results showed that although the perinatal health care providers' attitudes toward breast-feeding were very positive, there were many areas in which knowledge was incomplete and in which wide variation existed or incorrect advice given to breast-feeding mothers. The content of advice on breast-feeding was not always sound. Many thought that a wide range of illnesses were a contraindication to breast-feeding, nearly one in five thought that breast-feeding should follow a fixed schedule rather than the baby's needs, and only 54% thought that breast-feeding should be initiated immediately after delivery. Most of the providers did not seem to have the knowledge to cope with the common problem of insufficient breast-milk supply syndrome. Similarly, although support for the concept of rooming-in was strong, about one-third of respondents did not think the mother and infant should be together for the full 24 hr implied by true rooming-in. Fears about the possibility of increased risk of infection with rooming-in were expressed These and other misconceptions about rooming-in imply that a consistent, well-designed training program needs to be carried out in the modern health sector which will provide the necessary information to health care providers about this important aspect of early infant care.


Retinol-binding protein in human serum: conformational changes induced by retinoic acid binding.
Polyacrylamide gel electrophoresis and isoelectrofocusing followed by immunoblotting technique with an anti-human retinol-binding protein (RBP) serum were used to study holo-RBP and apo-RBP in human plasma. Three observations were made the technique allowed for the first time to directly and quantitatively analyse holo- and apo-RBP. Holo-RBP represented 97.86 +/- 0.78% and apo-RBP 1.94 +/- 0.73% of the total RBP. All-trans-retinoic acid (RA) was found to bind to apo-RBP and to significantly modify the tertiary structure of the protein; this raises the question of RBP involvement in the transport of RA. reconstitution of holo-RBP using apo-RBP from delipidized serum was achieved only after its incubation with natural all-trans-retinoids such as retinol, 3-dehydroretinol and retinoic acid but not with synthetic analogs of retinoic acid (13-cis-retinoic acid, TMMP, 13-cis-TMMP, TTNPB). It appears that RBP has a structure specificity for natural retinoids.


Calcium exchange in rabbit myocardium during and after hypoxia: role of sodium-calcium exchange.
Calcium uptake was measured using 47Ca2+ in the isolated and arterially perfused interventricular septum of the rabbit. Experiments were undertaken to determine whether calcium uptake on reoxygenation is linked to recovery of mechanical function and whether calcium uptake is through the sodium-calcium exchange mechanism. During substrate-free hypoxia for 45 min total tissue calcium remained unchanged but immediately upon reoxygenation there was a substantial net gain of calcium. Recovery of mechanical function upon reoxygenation was inversely related to the increase in tissue calcium. Activation of sodium-calcium exchange by perfusion with a low-sodium, zero-potassium, sucrose solution also increased tissue calcium and the relation to mechanical recovery was similar to that observed on reoxygenation. The sodium-calcium exchange mechanism was not affected by hypoxia and could be demonstrated during perfusion with a substrate-free hypoxic solution. Lithium (100 mM) substitution for sucrose prevented the calcium influx induced by a low-sodium and zero-potassium perfusate under normoxic conditions. Lithium substitution early during hypoxia or on reoxygenation did not affect the increase in myocardial calcium on reoxygenation. Amiloride (10(-4)M), presumed to inhibit sodium-hydrogen exchange during hypoxia, had no effect upon reoxygenation induced calcium uptake. It is concluded that the increase in calcium uptake that occurs on reoxygenation after a period of substrate-free hypoxia is related to mechanical recovery. Sodium-calcium exchange may contribute to calcium uptake on reoxygenation in this experimental model but is not the major mechanism.


Business risk and the health care entrepreneur.
This article addresses issues of business risk in the home health care industry. General concepts of risk are discussed, and the unique attributes affecting risk in the home health industry are identified and reviewed particularly as they affect market entry strategies, differentiation, innovation, and regulation.


Systematic review of endovascular versus laparoscopic extravascular stenting for treatment of nutcracker syndrome.
The aim of the present study was to assess the current strategies of endovascular and laparoscopic extravascular stenting for symptomatic compression of the left renal vein (LRV), most frequently between the aorta and superior mesenteric artery (nutcracker syndrome [NCS]). We performed a systematic review of all studies of endovascular and laparoscopic extravascular LRV stenting for NCS using the PubMed/MEDLINE, Scopus, Embase, Cochrane, Science Citation Index Expanded, Emerging Sources Citation Index, and Epistemonikos databases. Data were collected in accordance with the PRISMA (preferred reporting items for systematic reviews and meta-analysis) guidelines. The English, Spanish, and German language literature was searched from January 1, 1946 to February 9, 2022. The outcomes assessed included symptom resolution, hematuria resolution, and reintervention at follow-up. The search yielded 3498 reports. After removing the duplicates and those without the full text available, 1724 studies were screened. Of these, 11 studies were included in the present review. Of the 11 studies, 7 were on endovascular stenting and 4 on laparoscopic extravascular stenting; all 11 studies were retrospective, single-center case series. Of the 233 patients, 170 (80 women) had undergone endovascular stenting and 63 (9 women) had undergone extravascular stenting. The follow-up period varied from 1 to 60 months after endovascular stenting and 3 to 55 months after extravascular stenting. The symptoms had resolved in 76% (range, 50%-100%) after endovascular stenting and 83% (range, 71%-100%) after extravascular stenting. Hematuria had resolved in 86% (range, 60%-100%) after endovascular stenting and 89% (range, 77%-100%) after extravascular stenting. Of 185 patients, 9 had required reintervention after endovascular stenting and none after extravascular stenting. Endovascular and laparoscopic extravascular stenting are less invasive and, thus, more attractive treatment options that have been more recently developed for the management of NCS. The results from the present study have shown that symptom and hematuria resolution must be provided before they can be considered preferred management options for patients affected by NCS. Given the limited number of patients involved, no definitive conclusion could be drawn regarding the superiority of one technique compared with the other.


[Radiotherapy techniques of the mediastinum (author's transl)].
The principal techniques of mediastinal irradiation are discussed with reference to the need for rational space distribution of the dose, one reason being to respect healthy tissues (particularly the bone marrow); numerous examples are presented.


The French Randomized Optimal Stenting Trial: a prospective evaluation of provisional stenting guided by coronary velocity reserve and quantitative coronary angiography. F.R.O.S.T. Study Group.
We sought to make a prospective comparison of systematic stenting with provisional stenting guided by Doppler measurements of coronary velocity reserve and quantitative coronary angiography. Despite the increasing use of stents during percutaneous transluminal coronary angioplasty, it is unclear whether systematic stenting is superior to a strategy of provisional stenting in which stents are placed only in patients with unsatisfactory results or as a bail-out procedure. Two hundred fifty-one patients undergoing elective coronary angioplasty were randomly assigned either to provisional stenting (group 1, in which stenting was performed if postangioplasty coronary velocity reserve was <2.2 and/or residual stenosis > or =35% or as bail-out) or to systematic stenting (group 2). The primary end point was the six-month angiographic minimal lumen diameter (MLD). Major adverse cardiac events were secondary end points (death, acute myocardial infarction and target lesion revascularization). Stenting was performed in 48.4% of patients in group 1 and 100% of patients in group 2 (p<0.01). Six months after angioplasty, the MLD did not differ between groups (1.90+/-0.79 mm vs. 1.99+/-0.70 mm, p = 0.39), as was the rate of binary restenosis (27.1% vs. 21.4%, p = 0.37). Among patients with restenosis, 13/32 (40.6%) in group 1 but 100% (25/25) in group 2 had in-stent restenosis (p<0.01). Target lesion revascularization (15.1% vs. 14.4% in groups 1 and 2 respectively, p = 0.89) and major adverse cardiac events (15.1% vs. 16.0%, p = 0.85) were not significantly different. Systematic stenting does not provide superior angiographic results at six months as compared with provisional stenting.


CircRNA-mediated ceRNA mechanism in Osteoarthritis: Special emphasis on circRNAs in exosomes and the crosstalk of circRNAs and RNA methylation.
Osteoarthritis (OA) is an age-related joint disease with chronic inflammation, progressive articular cartilage destruction and subchondral bone sclerosis. CircRNAs (circRNAs) are a class of non-coding RNA with a circular structure that participate in a series of important pathophysiological processes of OA, especially its ceRNA mechanisms, and play an important role in OA. CircRNAs may be potential biomarkers for the diagnosis and prognosis of OA. Additionally, differentially expressed circRNAs were found in patients with OA, indicating that circRNAs are involved in the pathogenesis of OA. Experiments have shown that the intra-articular injection of modified circRNAs can effectively relieve OA. Exosomal circRNAs and methylated circRNAs also provide new ideas for the treatment of OA. Clarifying the important roles of circRNAs in OA will deepen people's understanding of the pathogenesis of OA. CircRNAs may be developed as new biomarkers or drug targets for the diagnosis of OA and provide new methods for the treatment of OA.


Surgical and endovascular intervention for infrainguinal vein graft stenosis.
The purpose of this study was to evaluate the stenosis-free patency of open repair (vein-patch angioplasty, interposition, jump grafting) and percutaneous transluminal balloon angioplasty (PTA) of 144 vein graft stenoses that were detected during duplex scan surveillance after infrainguinal vein bypass grafting. Patients who underwent revision of an infrainguinal vein bypass graft were analyzed for type of vein conduit, vascular laboratory findings leading to revision, repair techniques, assisted graft patency rate, procedure mortality rate, and restenosis of the repair site. The time of postoperative revision ranged from 1 day to 133 months (mean, 13 months). One hundred eighteen primary and 26 recurrent stenoses (peak systolic velocity, >300 cm/s) in 52 tibial and 35 popliteal vein bypass grafts were identified by means of duplex scanning. The repairs consisted of 77 open procedures (vein-patch angioplasty, 28; vein interposition, 33; jump graft, 9; primary repair, 3) and 67 PTAs. No patient died as a result of intervention. Cumulative assisted graft patency rate (life-table analysis) was 91% at 1 year and 80% at 3 years. At 2 years, cumulative assisted graft patency rate was comparable for saphenous vein grafts (reversed, 94%; in situ, 88%; nonreversed, 63%) and alternative vein grafts (89%). Stenosis-free patency rate at 2 years was identical (P =.55) for surgical intervention (63%) and endovascular intervention (63%) but varied with type of surgical revision (P =.04) and time of intervention (<4 months, 45%; >4 months, 71%; P =.006). The use of duplex scan-monitored PTA to treat focal stenoses (<2 cm) and late-appearing stenoses (>3 months) was associated with a stenosis-free patency rate that was 89% at 1 year. After intervention, the alternative vein bypass grafts necessitated twice the reinterventions per month of graft survival (P =.01). Bypass graft to the popliteal versus infrageniculate arteries, site of graft stenosis (vein conduit, anastomotic region), and repair of a primary versus a recurrent stenosis did not influence the outcome after intervention. The revision of duplex scan-detected vein graft stenosis with surgical or endovascular techniques was associated with an excellent patency rate, including when intervention on alternative vein conduits or treatment of restenosis was necessary. When PTA was selected on the basis of clinical and duplex scan selection criteria, the endovascular treatment of focal vein graft stenosis was effective, durable, and comparable with the surgical revision of more extensive lesions.


High-altitude-related disorders--Part II: prevention, special populations, and chronic medical conditions.
This second section of a 2-part review on high-altitude-related disorders focuses on strategies for prevention of high-altitude illness, identification of populations at increased risk for high-altitude illness, and effects of high altitude on selected chronic medical conditions. Practical aspects of advising and educating patients traveling to high altitude will be discussed, with special reference to pregnant women, infants and young children, healthy elders, and chronic medical conditions that may place persons at greater risk for high-altitude illness. The special concerns of pre-verbal children will be covered relative to the risks of high altitude for those too young to voice symptoms of illness and, thus, at-risk for potential serious consequences caused by delay in diagnosis and treatment.


PCPro: a clinically accessible, circulating lipid biomarker signature for poor-prognosis metastatic prostate cancer.
Using comprehensive plasma lipidomic profiling from men with metastatic castration-resistant prostate cancer (mCRPC), we have previously identified a poor-prognostic lipid profile associated with shorter overall survival (OS). In order to translate this biomarker into the clinic, these men must be identifiable via a clinically accessible, regulatory-compliant assay. A single regulatory-compliant liquid chromatography-mass spectrometry assay of candidate lipids was developed and tested on a mCRPC Discovery cohort of 105 men. Various risk-score Cox regression prognostic models of OS were built using the Discovery cohort. The model with the highest concordance index (PCPro) was chosen for validation and tested on an independent Validation cohort of 183 men. PCPro, the lipid biomarker, contains Cer(d18:1/18:0), Cer(d18:1/24:0), Cer(d18:1/24:1), triglycerides and total cholesterol. Within the Discovery and Validation cohorts, men who were PCPro positive had significantly shorter OS compared to those who were PCPro negative (Discovery: median OS 12.0 months vs 24.2 months, hazard ratio (HR) 3.75 [95% confidence interval (CI) 2.29-6.15], p < 0.001, Validation: median OS 13.0 months vs 25.7 months, HR = 2.13 [95% CI 1.46-3.12], p < 0.001). We have developed PCPro, a lipid biomarker assay capable of prospectively identifying men with mCRPC with a poor prognosis. Prospective clinical trials are required to determine if men who are PCPro positive will benefit from therapeutic agents targeting lipid metabolism.


Laser Doppler flux-metry as an experimental tool in laryngology.
Angiogenesis is a cascade-like mechanism, essential for the progression of solid tumours. Quantifying microvessel density is considered the gold standard for measuring baseline angiogenesis and indeed "the response to intervention". Applying immunohistochemical techniques to biopsies conventionally does this; however, it would be very helpful to have a less invasive, reliable method of quantifying blood flux, and hence angiogenesis. The aim of the study was to validate the use of a laser Doppler flux meter as an experimental tool in laryngology. The principle and technique of laser Doppler flux-metry is described. A validation study designed to estimate the reproducibility and ability to measure change was performed in the human larynx and in an animal model. Statistical advice was sought and analysis was performed using SPSS software. Our experiments show good reproducibility of laser Doppler measurements in the human and animal model (correlation coefficient 0.956 and 0.947, respectively, P=0.01). We have also shown that the laser Doppler flux meter is capable of detecting change when it is expected ( P<0.01). Utilising a standardised technique, laser Doppler flux-metry appears to be a reliable and reproducible method of estimating blood flux in the larynx.


The potential role of nanoyttria in alleviating oxidative stress biomarkers: Implications for Alzheimer's disease therapy.
Alzheimer's disease (AD) is a fatal neurodegenerative disease that requires immediate attention. Oxidative stress that leads to the generation of reactive oxygen species is a contributing factor to the disease progression by promoting synthesis and deposition of amyloid-β, the main hallmark protein in AD. It has been previously demonstrated that nanoyttria possesses antioxidant properties and can alleviate cellular oxidative injury in various toxicity and disease models. This review proposed that nanoyttria could be used for the treatment of AD. In this paper, the evidence on the antioxidant potential of nanoyttria is presented and its prospects on AD therapy are discussed.


Rhabdomyolysis in a Morbidly Obese Patient After Oral Cavity Free Flap Reconstruction.
Post-operative rhabdomyolysis is a rare but life-threatening condition. Less than ten cases have been described in the otolaryngic literature and, to our knowledge, no reports exist in the setting of oral reconstructive free tissue transfer. Case report presentation. We discuss the clinical course that lead to the diagnosis of rhabdomyolysis with special consideration to simultaneous microvascular anastomosis. Serial lab values were closely followed to prevent kidney injury while preventing flap congestion. Excessive fluid resuscitation in free flap reconstruction has been associated with increased post-operative complications and flap failure. We present a cautionary case to highlight the need for early diagnosis of rhabdomyolysis, a condition that may become more prevalent in the head and neck population as obesity, the main risk factor, continues to increase worldwide.


Treatment of hepatitis B and C following liver transplantation.
Advanced liver disease from hepatitis B virus (HBV) and hepatitis C virus (HCV) is the leading indication for orthotopic liver transplantation (OLT) worldwide. Our understanding of recurrent liver disease related to HBV and HCV in the setting of OLT has evolved rapidly in the past decade. Recurrent viral hepatitis may lead to graft failure, death, or the need for retransplantation. Until about a decade ago, HBV was considered a contraindication to OLT due to its frequent recurrence and development of associated liver disease. Medical therapy with hepatitis B immune globulin and nucleoside analogues has diminished the risk of HBV recurrence and led to improvement in patient and graft survival. Consequently, OLT is now considered to be the standard of care in patients with end-stage liver disease related to HBV. HCV recurrence after OLT is almost universal. Although short-term survival in patients undergoing OLT for HCV is similar to survival for those transplanted for other indications, recurrent HCV may have an impact on long-term patient and graft survival. A specific and effective therapy has not been defined for recurrent HCV following transplantation, but the combination of interferon and ribavirin appears promising. Optimal strategies to eradicate these viruses or to slow disease progression are continually being investigated in light of the disparity between supply and demand in a diminishing organ pool for OLT candidates.


Influence of high-fat diet on amine oxidase activity in white adipose tissue of mice prone or resistant to diet-induced obesity.
Decreased monoamine oxidase (MAO) activity has been observed in adipose tissue of obese patients. Since substrates of MAO and semicarbazide-sensitive amine oxidase (SSAO) can modify adipocyte metabolism, this work investigates whether changes in amine oxidase activity may occur during white adipose tissue (WAT) development. We evaluated MAO and SSAO activities in WAT of high-fat diet (HFD) and low-fat diet fed mice. To distinguish the effect of HFD on its own from the effect of fat mass enlargement, obesity-prone transgenic line of the FVBn strain lacking beta3-adrenergic receptors (AR) but expressing human beta3-AR and alpha2-AR (mbeta3-/-, hbeta3+/+, halpha2+/-) was compared to its obesity-resistant control (mbeta3-/-, hbeta3+/+). As already reported, the former mice became obese while the latter resisted to HFD. No significant change in SSAO or MAO activity was found in WAT of both strains after HFD when expressing oxidase activity per milligram of protein. However, when considering the overall capacity of the fat depots to oxidize tyramine or benzylamine, there was an increase in MAO and SSAO activity only in the enlarged WAT of HFD-induced obese mice. Therefore, the comparison of these models allowed to demonstrate that the higher amine oxidase capacity hold in enlarged fat stores of obese mice is more likely the consequence of increased fat cell number rather than the result of an increased expression of MAO or SSAO in the adipocyte.


Evaluation of therapeutic effect of Premna herbacea in diabetic rat and isoverbascoside against insulin resistance in L6 muscle cells through bioenergetics and stimulation of JNK and AKT/mTOR signaling cascade.
Premna herbacea Roxb., a perennial herb is well documented for its therapeutic uses among the traditional health care-givers of Assam, India. Scientific validation on the traditional use of the medicinal plant using modern technology may promote further research in health care. This study evaluates the therapeutic potential of methanolic extract of P. herbacea (MEPH) against type 2 diabetes mellitus (T2DM) and its phytochemical(s) in ameliorating insulin resistance (IR), thereby endorsing the plant bioactives as effective anti-hyperglycemic agents. The anti-diabetic potential of the plant extract was explored both in L6 muscle cells and high fructose high fat diet (HF-HFD) fed male Sprague Dawley (SD) rats. Bioactivity guided fractionation and isolation procedure yielded Verbascoside and Isoverbascoside (ISOVER) as bioactive and major phytochemicals in P. herbacea. The bioenergetics profile of bioactive ISOVER and its anti-hyperglycemic potential was validated in vitro by XFe24 analyzer, glucose uptake assay and intracellular ROS generation by flourometer, FACS and confocal microscopy. The potential of ISOVER was also checked by screening various protein markers via immunoblotting. MEPH enhanced glucose uptake in FFA-induced insulin resistant (IR) L6 muscle cells and decreased elevated blood glucose levels in HF-HFD fed rats. Isoverbascoside (ISOVER) was identified as most bioactive phytochemical for the first time from the plant in the Premna genus. ISOVER activated the protein kinase B/AMP-activated protein kinase signaling cascades and enhanced glucose uptake in IR-L6 muscle cells. ISOVER decreased the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK) and c-Jun N-terminal kinase (JNK) and increased that of mammalian target of rapamycin (mTOR), thereby attenuating IR. However, molecular docking revealed that ISOVER increases insulin sensitivity by targeting the JNK1 kinase as a competitive inhibitor rather than mTOR. These findings were further supported by the bioenergetics profile of ISOVER. This study for the first time depicts the functional properties of ISOVER, derived from Premna herbacea, in ameliorating IR. The phytochemical significantly altered IR with enhanced glucose uptake and inhibition of ROS through JNK-AKT/mTOR signaling which may pave the way for further research in T2DM therapeutics.


Trends of outcomes and healthcare utilization following orthopaedic procedures in adults with haemophilia: A 3-decade retrospective review.
Haemophilic arthropathy is a serious complication of haemophilia often requiring surgical intervention. It is unclear whether advances in comprehensive care are associated with a reduction in orthopaedic interventions and peri-procedural resource utilization. To determine temporal patterns of orthopaedic interventions in persons with haemophilia (PWH), and evaluate changes in healthcare utilization and outcomes. In this Canadian multicentre retrospective cohort study, adult PWH from Northern Alberta and British Columbia who underwent orthopaedic procedures (1990-2018) were included. Temporal changes in the type of procedures, length of stay (LOS), factor utilization and outcomes were examined. Sixty-five patients (78% haemophilia A) underwent 102 surgeries at a median age of 46.3. Of the 46 severe PWH, 28 (61%) were on prophylaxis at time of surgery. The proportion of total knee arthroplasties (TKA) declined over time (56% 1990-1999, 51% 2000-2009, 27% 2010-2018), with a concomitant rise in ankle arthrodesis (0% 1990-1999, 18% 2000-2009, 27% 2010-2018). Over time, PWH underwent orthopaedic procedures at an older age (P = .02). There was a significant reduction in perioperative factor VIII utilization (P = .003) and median LOS (P < .0001). Major bleeds, prosthetic joint infections and thrombosis were not observed in the last decade. In the last three decades, there was a decline in the proportion of TKA, likely reflecting the impact of widespread use of tertiary prophylaxis. However, ankle arthrodesis rates increased, suggesting that higher trough levels may be required to prevent ankle arthropathy. We observed a significant reduction in LOS and factor utilization, reflecting improvements in perioperative management.


Glutathione-stabilized Cu nanoclusters as fluorescent probes for sensing pH and vitamin B1.
Glutathione (GSH), playing roles as both a reducing reagent and protecting ligand, has been successfully employed for synthesizing Cu nanoclusters (CuNCs@GSH) on the basis of a simple and facile approach. The as-prepared CuNCs exhibited a fluorescence emission at 600nm with a quantum yield (QY) of approximately 3.6%. Subsequently, the CuNCs described here was employed as a broad-range pH sensor by virtue of the fluorescence intensity of CuNCs responding sensitively to pH fluctuating in a linear range of 4.0-12.0. Meanwhile, these prepared CuNCs were applied for detections of vitamin B1 (VB1) on the basis of positively charged VB1 neutralizing the negative surface charge of CuNCs, thus leading to the instability and aggregations of CuNCs, and further facilitating to quench their fluorescence. In addition, the proposed analytical method permitted detecting VB1 with a linear range of 2.0×10(-8)-1.0×10(-4) mol L(-1) as well as a detection limit of 4.6×10(-9) mol L(-1). Eventually, the practicability of this sensing approach was validated by assaying VB1 in human urine samples and pharmaceutical tablets, confirming its potential to broaden avenues for assaying VB1.


Dose-Response Relationship in Stereotactic Body Radiation Therapy for Hepatocellular Carcinoma: A Pooled Analysis of an Asian Liver Radiation Therapy Group Study.
Despite the worldwide implementation of stereotactic body radiation therapy (SBRT) for hepatocellular carcinoma (HCC), there is a lack of consensus guideline on prescription dose. Herein, this multinational study aimed to investigate the effects of the prescribed radiation dose on oncologic outcomes of SBRT for HCC. The multi-institutional retrospective cohort included 510 patients treated with SBRT between 2010 and 2016. All relevant clinical factors and factors related to SBRT were analyzed to evaluate freedom from local progression (FFLP) and overall survival (OS). Based on a biologically effective dose (BED) cutoff value of 100 Gy, 198 tumors were selected from each group in propensity score matching (PSM). Baseline characteristics in the BED <100 Gy group were unfavorable (Child-Pugh class B, 19%; advanced stage, 72%; median tumor size was 4 cm) compared with the BED ≥100 Gy group. With a median follow-up of 22 (interquartile range, 9.8-37.6) months, the 2-year FFLP and OS rates were 77% and 73%, respectively. Patients treated with a BED ≥100 Gy showed better rates of 2-year FFLP and OS than patients treated with a BED <100 Gy (FFLP, 89% vs 69%; OS, 80% vs 67%; P < .001). In the multivariable analysis before and after PSM, BED ≥100 Gy was identified as the main prognostic factor for both FFLP and OS (P < .01). Additionally, a dose-response relationship was observed between FFLP and BED (odds ratio, 0.92 per 5 Gy, P = .048). A BED ≥100 Gy was significantly associated with outcomes, and a dose-response relationship was observed between local tumor progression and BED. Given that SBRT is being increasingly used in HCC, detailed consensus guidelines regarding SBRT dose prescription should be established.


New insights from modelling and estimation of mass transfer parameters in fixed-bed adsorption of Bisphenol A onto carbon materials.
The removal of Bisphenol A, 2,2-bis (4-hydroxyphenyl) propane (BPA) in fixed-bed columns was investigated by breakthrough adsorption tests at different operation conditions and further prediction by a mathematical model to describe the adsorption-diffusion process onto two synthesized carbon porous materials. In this study, a xerogel (RFX) prepared by an optimized conventional sol-gel method and a lignin-based activated carbon (KLP) obtained via chemical activation were used in batch and fixed-bed adsorption experiments. The materials were fully characterized and their adsorptive properties were compared to those obtained with a commercial activated carbon (F400). RFX and KLP materials reached the equilibrium adsorption in only 24 h, whereas F400 activated carbon required 48 h. In addition, F400 and KLP adsorbents showed higher equilibrium adsorption capacity values (qe = 0.40 and 0.22 kg/kg, for F400 and KLP, respectively) than that obtained for the xerogel (qe = 0.08 kg/kg). Both synthesized carbon-adsorbents were studied in fixed-bed adsorption tests, exploring the effect of the operation conditions, e.g., initial BPA concentration (0.005-0.04 kg/m3), weight of adsorbent (0.01-0.05 g) and volumetric flow rate (0.2 to 1.0 mL/min), on the adsorption performance of the column. All the tested adsorption columns reached the equilibrium in a very short time, due to the efficient dimensionless of the bed. Additionally, the regeneration of the exhausted adsorbent was studied, achieving the total reuse of the solids after three consecutive cycles using methanol as regeneration agent. Finally, a mathematical model based on mass conservation equations was proposed, allowing to efficiently fit the experimental BPA breakthrough curves and estimate the external and adsorbed-phase mass transfer coefficients with a high accuracy.


[Portal anticoagulation in preventing thrombosis after porta-azygous devascularization for portal hypertension].
To study the prevention method for thrombosis after porta-azygous devascularization for portal hypertension. 71 patients with portal hypertension due to cirrhosis were divided into group A (36 patients) and group B (35 patients). In group A, a anticoagulation tube was inserted via splenic vein branch during shunt and anticoagulation therapy was given after operation. All the patients received color Doppler examination 3 months after operation. No thrombosis occurred in the group A and thrombosis occurred in 13 patients (37.14%) in the group B. Portal anticoagulation therapy is the effective in preventing portal thrombosis after porta-azygous devascularization for portal hypertension.


PTH stimulates the proliferation of TE-85 human osteosarcoma cells by a mechanism not involving either increased cAMP or increased secretion of IGF-I, IGF-II or TGF beta.
Injections of parathyroid hormone (PTH) result in increased bone formation in several species. Work in our laboratory and others has shown a stimulation of bone cell proliferation and growth factor production by PTH. Our purpose was to study the effects of PTH on a human bone cell line using TE-85 human osteosarcoma cells as a model. After 24 h treatment, PTH caused an increase in cell proliferation as measured by cell counts and [3H]-thymidine incorporation. Proliferation was not inhibited by an anti-transforming growth factor beta (TGF beta) antibody which could abolish stimulation by exogenous TGF beta. PTH did not stimulate cAMP production, alkaline phosphatase activity or production of insulin-like growth factors I or II (IGF-I or IGF-II) in TE-85 cells. Although basal TE-85 proliferation was slowed by incubation with the calcium channel blocking agent verapamil, PTH still caused an increase in growth rate. We conclude that PTH directly stimulates TE-85 proliferation via a mechanism not involving increased adenylate cyclase activity or increased secretion of IGF-I, IGF-II or TGF beta and may stimulate bone formation in vivo by activating some other mitogenic signal to increase bone cell proliferation.


Evidence against increased glomerular pressure initiating diabetic nephropathy.
Studies were carried out to determine whether exaggerated glomerular hydraulic pressure (PG) initiates the development of glomerular pathology and proteinuria in insulin-dependent diabetic rats. Normotensive (WKY) and hypertensive rats (SHR) were made diabetic by streptozotocin injection. One group of SHR diabetic rats was treated with antihypertensive drugs to reduce blood pressure. One week after onset of diabetes, micropuncture determinations of PG, measured by stopped-flow technique, revealed that PG was higher in WKY diabetics than in non-diabetic WKY controls, and that PG was even higher in SHR diabetics (P less than 0.05). Similarly prepared groups of animals were followed for six months, approximately one fifth to one third of the expected life span of these rats. Tail systolic blood-pressure measurements documented continuous severe systolic-hypertension in SHR diabetics, normal pressure in the WKY diabetics and hypotension in the SHR diabetics treated continuously with antihypertensive drugs. Urinary protein excretion, measured monthly, was statistically the same in all groups, with no evidence of a progressive rise in the SHR diabetics. PG measured in two rats from each group after four months of diabetes was similar to values found after one week of diabetes. Semiquantitative histologic scoring of glomerular mesangial expansion after six months of diabetes failed to demonstrate any significant difference between the normotensive WKY diabetics and the hypertensive SHR diabetics. These observations suggest that elevated PG does not in itself initiate glomerular pathology or proteinuria in diabetes.(ABSTRACT TRUNCATED AT 250 WORDS)


Effective plant-endophyte interplay can improve the cadmium hyperaccumulation in Brachiaria mutica.
Soil contamination due to cadmium (Cd) is a ubiquitous environmental problem for which inexpensive remediation alternatives are required. Phytoaccumulation, the use of plants to extract and accumulate heavy metals from the contaminated environment, is such an alternative. In this study, we aimed at establishing effective plant-bacteria interplay between Brachiaria mutica and Cd-resistant endophytic bacteria eventually leading to improved phytoremediation. B. mutica was grown in a Cd-contaminated soil and inoculated with three Cd-tolerant endophytic bacteria individually as well as in combination. Plant physiological parameters, biomass production, bacterial colonization, and Cd-accumulation were observed at four different Cd exposures, i.e., 100, 200, 400 and 1000 mg kg-1 of soil. The combined application of endophytic bacteria was more effective as compared to their individual applications at all concentrations. Nevertheless, highest performance of consortium was seen at 100 mg Cd kg-1 of soil, i.e., root length was enhanced by 46%, shoot length by 62%, chlorophyll content by 40%, and dry biomass by 64%; which was reduced with the increase in Cd concentration. The bacterial population was highest in the root interior followed by rhizosphere and shoot interior. Concomitantly, plants inoculated with bacterial consortium displayed more Cd-accumulation in the roots (95%), shoots (55%), and leaves (44%). Higher values of BCFroot (> 1), and lower values for BCFshoot and TF (< 1) indicates capability of B. mutica to accumulate high amounts of Cd in the roots as compared to the aerial parts. The present study concludes that plant-endophyte interplay could be a sustainable and effective strategy for Cd removal from the contaminated soils.


Adsorption of ionic surfactants at the air-water interface: The gap between theory and experiment.
We review the experimental and theoretical results for the adsorption and structure of ionic surfactants at the air-liquid interface. The results show that ionic surfactants form thick adsorption layers at the interfacial region. We also review several adsorption models for ionic surfactants, which become increasingly complex as they capture the many features of adsorption layers. However, the adsorption layer structures determined by experiments and the structures predicted by models do not match because most models assume very thin adsorption layers. We show the discrepancies between measured and predicted surface properties and provide several explanations. We conclude that the mismatch in the adsorption layer structure provided by experiments and the structure provided by adsorption models is the main reason for the discrepancies in the surface excess and the surface potential.


Neurological infections after neuraxial anesthesia.
Infection is the commonest cause of serious neurologic sequelae of neuraxial anesthesia. The incidence depends on operator skill and patient population. Meningitis, a complication of dural puncture, is usually caused by viridans streptococci. The risk factors are dural puncture during labor, no mask and poor aseptic technique, vaginal infection and bacteremia. Epidural abscess is a complication of epidural catheterization, route of entry the catheter track and the organism usually the staphylococcus. Principal risk factors are prolonged catheterization, poor aseptic technique and traumatic insertion. Prevention includes wearing a mask, using a full sterile technique, avoiding prolonged catheterization and prescribing antibiotics in a high-risk situation.


Diagnostic values of leukocyte, neutrophil, and neutrophil to lymphocyte ratio in distinguishing central facial paralysis from peripheral facial paralysis.
Misdiagnosis of central facial palsy (CFP) as peripheral facial palsy (PFP) can lead to serious consequences clinically. It is unknown whether the leukocyte counts (leukocyte), neutrophil counts (neutrophil), and neutrophil to lymphocyte ratio (NLR) can be used to distinguish CFP from PFP. Of the total 152 patients admitted for acute facial paralysis, 76 CFP patients (CFP group) caused by acute ischemic stroke (AIS) and 76 PFP cases (PFP group) without AIS were enrolled in this retrospective study. The levels of blood leukocyte, neutrophil, lymphocyte, platelet counts (platelet), NLR, and platelet to lymphocyte ratio (PLR) before or upon admission were recorded and compared between the two groups. The student t-test was adopted for comparison of the mean. Model discrimination was evaluated using the area under the receiver operating characteristic curve (AUC). Comparison of AUC was performed using the Z-test. Compared with PFP group, the levels of leukocyte, neutrophil, and NLR were significantly increased in CFP group (all p < 0.01), and there were still significantly statistical differences (all p < 0.01), even after adjusting for age, gender, and past medical history, while no significantly statistical differences of lymphocyte, platelet, and PLR were found between CFP and PFP (all p > 0.05); furthermore, the AUC in distinguishing CFP from PFP were 0.629, 0.671, and 0.657 for leukocyte, neutrophil, and NLR, respectively, and no significant difference of AUC was observed among leukocyte, neutrophil, and NLR (p > 0.05); finally, the cutoff values (specificity, sensitivity, and Youden index) in distinguishing CFP from PFP were 7.08 × 109/L (65.79%, 57.89%, 0.237) for leukocyte, 4.90 × 109/L (73.68%, 60.53%, 0.342) for neutrophil, and 2.88 (72.37%, 55.26%, 0.276) for NLR, respectively. As easy-to-obtain and inexpensive inflammatory biomarkers, leukocyte, neutrophil, and NLR could demonstrate diagnostic values in distinguishing between CFP and PFP.


Abnormally High Blood Acetaldehyde Concentrations Suggest Potential Postmortem Ethanol Generation.
Ethanol is one of the most commonly used and abused substances worldwide. Identifying whether the source of ethanol detected in corpses is antemortem ingestion or postmortem generation is especially important for determining the cause of death, which remains a vibrant field of research. During the synthesis of ethanol in the putrefaction process of corpses, other small molecules such as acetaldehyde and n-propanol could also be produced. According to our prospective statistical analysis based on authentic samples from forensic cases, it is rational to suspect ethanol generation after death when the concentration of acetaldehyde detected in blood exceeds 0.014 g/dL. Through in vitro simulation experiments, in addition to confirming that ethyl glucuronide and ethyl sulfate are the reliable biomarkers of antemortem ingestion of ethanol, we propose that acetaldehyde is far more sensitive than n-propanol as a potential marker in the blood of corpses for postmortem ethanol formation.


A study of multiple causes of death in California: 1955 and 1980.
Multiple cause of death patterns in California for 1980 were compared to a similar study of deaths conducted in 1955. Primary underlying causes of death changed, mainly reflecting the emergence of respiratory cancer as a major cause of death in 1980. The number of causes reported per death increased from 1955 to 1980, in all age and sex groups. Diseases of the arteries and pneumonia, which are among the most common underlying causes of death, appeared more often on death certificates in both 1955 and 1980 as contributing causes than as underlying the death. Diabetes was studied in detail in the 1955 report, and comparisons were made in 1980 to show increases in the proportions of deaths with this disease and corresponding increases in its prevalence among the living population from the National Health Survey. Multiple cause of death data can provide further information on the prevalence of a fatal disease in a population and its relative role in contributing to mortality, and can also provide new information on diseases that contribute to deaths, which was not previously available in population-based studies of single cause of death.


Human serum IgE against two major allergens from cow hair and dander. Determination of affinity and quantity of antigen-specific IgE.
The quantity and quality of antigen-specific IgE against purified major allergens from cow hair and dander was measured by a double-antibody radioimmunoassay. In serum from patients with allergic asthma to cow the affinity constant varied from 1 to 150 X 10(9) l/mol and the number of antibody combining sites from 0.01 to 0.4 X 10-9 mol/l serum for each antigen.


Catecholamines inhibit leukotriene formation and decrease leukotriene/prostaglandin ratio.
Adrenaline, noradrenaline, isoprenaline, and to a lesser extent dopamine inhibit the release of leukotriene (LT) B2 from calcium ionophore-stimulated human polymorphonuclear leukocytes, while the release of prostaglandin (PG) E2 is proportionally elevated. The inactivity of salbutamol, a noncatechol adrenergic beta 2-receptor agonist, and the inability of propranolol to antagonize the effects of adrenaline, suggest the mediation through beta-receptor independent mechanisms. Neither are alpha-1-receptors involved, as prazosin, a specific antagonist, fails to inhibit the reaction. As the principles for biochemical regulation of LT- and PG-production are met by catecholamines in several tissues, the mechanism is considered to be of general physiological importance. Catecholamines may function as coenzymes/antioxidants which, by altering the redox state of the enzyme iron or heme, decrease the LT/PG ratio thus protecting the organism against tissue anaphylaxis and other LT-related pathophysiology.


Effect of dexamethasone on serum protein extravasation in experimental brain infarcts of monkey: an immunohistochemical study.
Experimental brain infarcts were produced in 12 adult baboons (Papio cynocephalus) by transorbital permanent clipping of the left middle cerebral artery. One group (seven monkeys) received daily injections of 1 mg/kg dexamethasone, starting 1 h after vascular occlusion and continuing till the end of the experiment. Another group (five monkeys) was not treated. One week after vascular occlusion the volume of infarcts and peri-infarct edema was estimated morphometrically on histological sections, using Masson's trichrome stain and the peroxidase-anti-peroxidase (PAP) technique for visualization of serum protein extravasation. In the untreated animals the average volume of infarct was 6.57 +/- 4.23% (mean +/- SD) and the volume of edema 7.83 +/- 2.93% of ipsilateral hemisphere. In the treated animals the infarct volume was not different (7.95 +/- 3.00%), but the volume of peri-infarct edema was significantly lower (2.82 +/- 3.06%, p less than 0.05). The results obtained indicate that dexamethasone treatment reduces the development of peri-infarct edema but does not influence the size of infarcts.


Borrelia multiplex: a bead-based multiplex assay for the simultaneous detection of Borrelia specific IgG/IgM class antibodies.
Lyme borreliosis (LB) is the most common tick-borne infectious disease in the northern hemisphere. The diagnosis of LB is usually made by clinical symptoms and subsequently supported by serology. In Europe, a two-step testing consisting of an enzyme-linked immunosorbent assay (ELISA) and an immunoblot is recommended. However, due to the low sensitivity of the currently available tests, antibody detection is sometimes inaccurate, especially in the early phase of infection, leading to underdiagnoses. To improve upon Borrelia diagnostics, we developed a multiplex Borrelia immunoassay (Borrelia multiplex), which utilizes the new INTELLIFLEX platform, enabling the simultaneous dual detection of IgG and IgM antibodies, saving further time and reducing the biosample material requirement. In order to enable correct classification, the Borrelia multiplex contains eight antigens from the five human pathogenic Borrelia species known in Europe. Six antigens are known to mainly induce an IgG response and two antigens are predominant for an IgM response. To validate the assay, we compared the Borrelia multiplex to a commercial bead-based immunoassay resulting in an overall assay sensitivity of 93.7% (95% CI 84.8-97.5%) and a specificity of 96.5% (95%CI 93.5-98.1%). To confirm the calculated sensitivity and specificity, a comparison with a conventional 2-step diagnostics was performed. With this comparison, we obtained a sensitivity of 95.2% (95% CI 84.2-99.2%) and a specificity of 93.0% (95% CI 90.6-94.7%). Borrelia multiplex is a highly reproducible cost- and time-effective assay that enables the profiling of antibodies against several individual antigens simultaneously.


Deficiencies in paediatric research applications delaying ethics committee approval.
A clinical research application must be submitted for approval by a competent ethics committee (EC) before a study can be executed. There is very limited information on how such submissions could be optimised, especially regarding research in children and adolescents, which requires particular caution and age-adapted patient information. We assessed all research applications from the University Children's Hospital Zurich submitted to the EC of the Canton of Zurich in 2014–2015, i.e., in the first two years after Switzerland's new Human Research Act came into effect. Moreover, we validated our findings by assessing a randomly selected sample of applications from the same hospital in 2018–2019. We assessed a total of 86 applications from 2014–2015, originating from 29 departments and sub-specialties. The EC judged that it was not responsible for three applications and declined an assessment for another three because the studies had already been conducted. Thus, we included 80 applications in the present analysis (18 clinical trials, 52 research projects, 10 further use projects). Applicants withdrew four applications before the EC's final decision and the EC rejected two after assessment. The EC had objections in 46 (62%) of the remaining 74 applications. Formal, including formal legal, objections (n = 503) and legal objections (n = 287) accounted for the vast majority of objections. There were also 71 ethical and 82 scientific objections. The most frequent formal and formal legal objections were incomplete or missing age-adapted patient information (49%) and incorrect templates for informed consent and signature forms (46%). A review of the 20 randomly selected applications from 2018–2019 confirmed that four out of the five most frequent deficiencies relating to informed consent were identical to those observed in the 2014–2015 applications. Careful preparation of submission documents by the investigators and close adherence to formal and legal requirements have the potential to considerably optimise and expedite the EC review process, and thus the commencement of the clinical research.   &nbsp.


Genome-wide analysis of RNA-chromatin interactions in lizards as a mean for functional lncRNA identification.
Long non-coding RNAs (lncRNAs) are defined as transcribed molecules longer than 200 nucleotides with little to no protein-coding potential. LncRNAs can regulate gene expression of nearby genes (cis-acting) or genes located on other chromosomes (trans-acting). Several methodologies have been developed to capture lncRNAs associated with chromatin at a genome-wide level. Analysis of RNA-DNA contacts can be combined with epigenetic and RNA-seq data to define potential lncRNAs involved in the regulation of gene expression. We performed Chromatin Associated RNA sequencing (ChAR-seq) in Anolis carolinensis to obtain the genome-wide map of the associations that RNA molecules have with chromatin. We analyzed the frequency of DNA contacts for different classes of RNAs and were able to define cis- and trans-acting lncRNAs. We integrated the ChAR-seq map of RNA-DNA contacts with epigenetic data for the acetylation of lysine 16 on histone H4 (H4K16ac), a mark connected to actively transcribed chromatin in lizards. We successfully identified three trans-acting lncRNAs significantly associated with the H4K16ac signal, which are likely involved in the regulation of gene expression in A. carolinensis. We show that the ChAR-seq method is a powerful tool to explore the RNA-DNA map of interactions. Moreover, in combination with epigenetic data, ChAR-seq can be applied in non-model species to establish potential roles for predicted lncRNAs that lack functional annotations.


A comparison of real-time scanning with conventional static B-mode scanning.
To determine the relative value of linear-array real-time scanning compared with static B-mode scanning, a prospective study was carried out. The results indicate that real-time scanning is better. Advantages and disadvantages of each method are discussed.


[Risk factors of hypoparathyroidism following total or near total thyroidectomy].
To study the risk factors for postoperative hypoparathyroidism or hypocalcemia. Totally 414 patients with thyroid diseases who underwent total or near total thyroidectomy at Department of Head and Neck Surgery, Peking University Cancer Hospital and Institute from June 2007 to June 2014 were studied retrospectively. There were 119 male and 295 female patients with a median age of 47 years. The clinical and pathological features that related to post-operative hypoparathyroidism were studied by χ(2) test and multivariate Logistic regression analysis. Of the 414 patients, 36.2% developed transient hypocalcemia, 36.5% developed transient hypoparathyroidism, 2.2% developed permanent hypoparathyroidism. In regression analysis, unilateral or bilateral center lymph node dissection were associated with mild transient hypocalcemia after surgery (OR=2.366, P=0.022; OR=5.216, P=0.000); unilateral or bilateral center lymph node dissection as well as surgical options were significant risk factors for severe transient hypocalcemia (OR=4.029, P=0.001; OR=8.384, P=0.000; OR=2.073, P=0.017) and hypoparathyroidism (OR=1.755, P=0.040; OR=4.144, P=0.000; OR=2.287, P=0.000). The parathyroid hormone concentration on postoperative day 1 was an independent risk factor for permanent hypoparathyroidism (OR=2.011, P=0.014). The concentration of parathyroid hormone threshold <5.28 ng/L was a predictor to permanent hypoparathyroidism with accuracy of 95.0%. Bilateral center lymph node dissection is a risk factor of permanent hypoparathyroidism in patients received total thyroidectomy should be taken thoughtfully. The parathyroid hormone concentration on postoperative day 1 provides better prediction for persistent hypoparathyroidism.


Patient-centric drug product development: Acceptability across patient populations - Science and evidence.
The 6th APV (Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnologie e.V., The International Association for Pharmaceutical Technology) Winter Conference took place in Salzburg (Austria) from January 19-20, 2023. This conference was dedicated to advance patient-centric drug development across all dosage forms, indications and patient populations and was organized by the APV PaCeMe IN Task Force. The topic was chosen due to emerging evidence and increasing regulatory requirements to consider patient needs and capabilities in drug product development. It is well acknowledged that acceptability of a drug product and its dosage form is a fundamental aspect of patient centric drug product design which can directly impact adherence and intended use, hence effectiveness and safety. Despite the requirement to proof acceptability within the drug development program, respective methods to determine and compare the degree of acceptability of different dosage forms and drug product designs are still limited.


Do new cement-based mortars pose a significant threat to the aquatic environment?
Nowadays, the use of multi-functional mortars has increased significantly, with interesting applications in the sustainable construction. In the environment, the cement-based materials are subjected to leaching, so the assessment of potential adverse effects upon aquatic ecosystem is necessary. This study focuses on the evaluation of the ecotoxicological threat and of a new type of cement-based mortar (CPM-D) and its raw materials leachates. A screening risk assessment were performed by Hazard Quotient methods. The ecotoxicological effects were investigated by a test battery with bacteria, crustacean, and algae. Two different procedures, Toxicity test Battery Index (TBI) and Toxicity Classification System (TCS), to obtain a single value for toxicity rank were used. Raw materials showed the highest metal mobility and in particular, for Cu, Cd and V potential hazard was evidenced. Leachate toxicity assessment evidenced the highest effects linked to cement and glass while the mortar showed the lowest ecotoxicological risk. TBI procedure allows a finer classification of effect linked to materials with respect to TCS which is based on worst case approach. A safe by design approach taking into account the potential and the effective hazard of the raw materials and of their combinations could allow to achieve sustainable formulations for building materials.


Relationship between systemic immune inflammation index and amputation in patients with diabetic foot ulcer.
The systemic immune inflammation index (SII) is a cost-effective biomarker calculated by lymphocyte, neutrophil and platelet counts and is currently being studied in various diseases. Since there is no study examining the relationship between SII and diabetic foot ulcers (DFU) in the literature, our aim was to investigate the relationship between SII and amputation rate in DFU. Type 2 DM 511 patients with DFU were screened from 2017 to 2021. Laboratory data obtained on the first day of hospitalization were considered. Platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR) and SII were calculated from routine blood count. Participants were divided into two groups as amputation (Group 1) and non-amputation (Group 2). Amputation rate was 18.8%. The A1c (8.80 (3.26) % vs. 9.52 (3.10) %, p = 0.007) and HGB (10.17 ± 2.16 g/dL vs. 12.05 ± 2.20 g/dL, p < 0.001) levels, and lymphocyte count (1.81 (1.16) vs. 2.05 (1.11), p = 0.015) were significantly lower in Group 1 than Group 2. The counts of WBC (14.01 (9.16) × 109/L vs. 10.41 (5.82) × 109/L), PLT (393.35 (196.98) × 109/L vs. 312.05 (141.33) × 109/L), neutrophil (11.52 (8.75) × 109/L vs. 6.93 (5.96) × 109/L), PLR (226.04 (159.24) × 109/L vs. 153.12 (101.91) × 109/L), NLR (6.64 (6.93) vs. 3.34 (3.99)) and SII (2505.86 (3957.47) × 109/L vs. 1092.50 (1476.08) × 109/L), and the levels of CRP (14.12 (12.66) mg/dL vs. 3.86 (12.63) mg/dL) and ESR (87.50 (50.50) mm/h vs. 63.00 (57.25) mm/h) were significantly higher in Group 1 than Group 2 (all p < 0.001). AUC of ROC analysis of PLR was 0.666 (95% CI, 0.604-0.728), NLR was 0.695 (95% CI, 0.638-0.752) and SII was 0.716 (95% CI, 0.661-0.772) for the predicting of amputation and the SII had the best AUC with 67.4% sensitivity and 63.3%specificty. SII is a cost-effective and readily available marker, but alone may not be sufficient to predict the risk of amputation in DFU. In our results, the predictive role of SII alone or with other markers for future DFU and its role in predicting other chronic diabetic complications will be evaluated in extensive studies.


Effect of immunization with lipid associated polysaccharide antigen and anti CD-2 antibodies on class II MHC expression and cellular immune response in BALB/C mice infected with Leishmania donovani.
In a bid to characterize the antigens and immunization mechanisms which may be used to produce a protective response against L. donovani, role of lipid associated polysaccharide (LPS) antigen and whole antigen was evaluated. BALB/C mice were immunized with whole or LPS antigen in combination with one of three putative adjuvents (anti CD-2 antibody/FIA/0.85% Saline). LPS antigen emulsified in anti CD-2 antibody was found to induce significant antibodies in mice on day 28 against challenge with lethal dose of L. donovani. Immunoprophylactic properties of LPS and whole antigen was investigated on day 40 through cytokine elicitation (IL-2), MIF) in culture supernatants of spleen cells, but before that MHC-II expressed on macrophage was studied. The LPS antigen in combination with anti CD-2 antibody was found to be most immuno-reactive inducing higher MHC-II expression on macrophages which was associated with substantial rise in the level of MIF and IL-2. It coincided with decline in antibody titre in 100% mice immunized with LPS antigen while Leishmania injected as whole antigen failed to induce specific macrophage and T-cell response with all the above formulations. We surmise from our data that lipid associated polysaccharide antigen linked to anti CD-2 antibody has potential for eliciting protective immunity against Leishmania.


Longitudinal microSPECt/CT imaging and pharmacokinetics of synthetic luteinizing hormone-releasing hormone (LHRH) vaccine in rats.
Luteinizing hormone-releasing hormone (LHRH)-derived decapeptide-based vaccines have been used in studies of immunocastration and immunotherapy of prostate cancer, but no image data are available on the kinetics of vaccines post injection (p.i.). Therefore, an 131I radiolabeled LHRH-derived immunogen was developed to visualize and evaluate the retention of LHRH-derived vaccines in rats. The LHRH immunogens, which contained equal moles of 131I-p607E, p667 and p500, were formulated with an equal volume of an adjuvant, Montanide ISA50. MicroSPECT/CT imaging was performed to visualize the retention of the radiolabeled immunogen up to 30 days after intramuscular inoculation of 25 microg immunogens. The pharmacokinetics, distribution and excretion were also evaluated. The radiochemical purity of 131I-p607E was 97.85+/-2.12%. The longitudinal microSPECT/CT imaging revealed that most 131I-p607E was retained at the injected muscle site until 30 days p.i.. Biodistribution showed that 34.56+/-4.27% of radioactivity remained at the injected muscle site at 28 days p.i.. The cumulative radioactivity excreted via urine was 30.02+/-3.82% up to day 28 p.i.. The elimination half-life (t1/2), Tmax and Cmax were 158.67 h, 24 h, and 0.026 percentage of injected dose per gram (%ID/g), respectively. The LHRH immunogen, 131I-p607E, was mainly retained at the intramuscular injection site during the whole study period. The microSPECTICT imaging modality can be used to monitor the location and distribution of the LHRH immunogen, 131I-p607E, in a rat model.