The importance of early identification of infusion-related reactions to monoclonal antibodies.
Monoclonal antibodies constitute important and useful tools in clinical practice and biotechnology for diagnosing and treating infectious, inflammatory, immunological and neoplastic diseases. This article reviews evidence on the different acute adverse effects of monoclonal antibodies, specifically infusion-related reactions (IRRs), and on the measures that should be taken before and during crises. A literature search using key terms relating to IRRs produced by monoclonal antibodies was undertaken to generate a comprehensive narrative review of the information available. Immunomodulatory monoclonal antibodies may produce IRRs and hypersensitivity-related reactions. Strategies to avoid or minimize the appearance of IRRs depend on the monoclonal antibody and type of patient and reaction (pre-medication, slowing infusion rates, infusion interruption or desensitization, etc.). Considering the great number of available monoclonal antibodies in current practice and those which will soon be authorized, it is mandatory to have clear guidelines that can give support to practitioners and nurses to help them respond quickly and safely to the different IRRs related to the use of these therapeutic drugs.


Use of real-time PCR with propidium monoazide for enumeration of viable Escherichia coli in anaerobic digestion.
A combination of propidium monoazide (PMA) with real-time quantitative polymerase chain reaction (PMA-qPCR) was optimized to enumerate only viable Escherichia coli in anaerobic digestion processes. Repeating the PMA treatment twice and a final concentration of 100 μM resulted in an effective exclusion of DNA from heat-treated E. coli cells. In three anaerobic digestion processes, real-time PCR, PMA-qPCR, and the most probable number method (MPN) were used to estimate the numbers of total, viable, and culturable E. coli cells, respectively. Culturable concentrations of fecal coliforms were also measured by the membrane filter method. For thermophilic digestion, the reductions in total and viable E. coli cells from the digester influent to the effluent were significantly lower than those in culturable cells and fecal coliforms by two to four orders of magnitude. For mesophilic digestion, the differences in the reductions in E. coli and fecal coliforms counts were less than two orders of magnitude. Based on the measurements of viable E. coli determined by the PMA-qPCR method, the microbial quality of digester effluents was discussed for agricultural application, and pasteurization after anaerobic digestion was suggested for the destruction of viable pathogens.


Polymer microring resonators for high-sensitivity and wideband photoacoustic imaging.
Polymer microring resonators have been exploited for high-sensitivity and wideband photoacoustic imaging. To demonstrate high-sensitivity ultrasound detection, highfrequency photoacoustic imaging of a 49-microm-diameter black bead at an imaging depth of 5 mm was imaged photoacoustically using a synthetic 2-D array with 249 elements and a low laser fluence of 0.35 mJ/cm(2). A bandpass filter with a center frequency of 28 MHz and a bandwidth of 16 MHz was applied to all element data but without signal averaging, and a signal-to-noise ratio of 16.4 dB was obtained. A wideband detector response is essential for imaging reconstruction of multiscale objects, e.g., various sizes of tissues, by using a range of characteristic acoustic wavelengths. A simulation of photoacoustic tomography of beads shows that objects with their boundaries characteristic of high spatial frequencies and the inner structure primarily of low spatial frequency components can be faithfully reconstructed using such a detector. Photoacoustic tomography experiments of 49- and 301-microm-diameter beads were presented. A high resolution of 12.5 microm was obtained. The boundary of a 301-microm bead was imaged clearly. The results demonstrated that the high sensitivity and broadband response of polymer microring resonators have potential for high resolution and high-fidelity photoacoustic imaging.


A novel inhibitor of fatty acid synthase shows activity against HER2+ breast cancer xenografts and is active in anti-HER2 drug-resistant cell lines.
Inhibiting the enzyme Fatty Acid Synthase (FASN) leads to apoptosis of breast carcinoma cells, and this is linked to human epidermal growth factor receptor 2 (HER2) signaling pathways in models of simultaneous expression of FASN and HER2. In a xenograft model of breast carcinoma cells that are FASN+ and HER2+, we have characterised the anticancer activity and the toxicity profile of G28UCM, the lead compound of a novel family of synthetic FASN inhibitors. In vitro, we analysed the cellular and molecular interactions of combining G28UCM with anti-HER drugs. Finally, we tested the cytotoxic ability of G28UCM on breast cancer cells resistant to trastuzumab or lapatinib, that we developed in our laboratory. In vivo, G28UCM reduced the size of 5 out of 14 established xenografts. In the responding tumours, we observed inhibition of FASN activity, cleavage of poly-ADPribose polymerase (PARP) and a decrease of p-HER2, p- protein kinase B (AKT) and p-ERK1/2, which were not observed in the nonresponding tumours. In the G28UCM-treated animals, no significant toxicities occurred, and weight loss was not observed. In vitro, G28UCM showed marked synergistic interactions with trastuzumab, lapatinib, erlotinib or gefitinib (but not with cetuximab), which correlated with increases in apoptosis and with decreases in the activation of HER2, extracellular signal-regulated kinase (ERK)1/2 and AKT. In trastuzumab-resistant and in lapatinib-resistant breast cancer cells, in which trastuzumab and lapatinib were not effective, G28UCM retained the anticancer activity observed in the parental cells. G28UCM inhibits fatty acid synthase (FASN) activity and the growth of breast carcinoma xenografts in vivo, and is active in cells with acquired resistance to anti-HER2 drugs, which make it a candidate for further pre-clinical development.


The improvement of dynamic characteristics of ultrasonic therapeutic transducers using fine-grain PZT-based piezoceramics.
Optimizing the dynamic resonant characteristics of ultrasonic therapeutic transducers depends most importantly on fine-grain piezoceramics with good resonant properties. In this paper, we prepare and compare modified Pb(0.99)Sr(0.01)[0.03(Mn(1/3)Nb(2/3))-0.97(Zr(0.51)Ti(0.49))]O(3) piezoceramics with 0.1 wt% CaCO(3) and 0.8 wt% PbO additives (PMZT3) synthesized by B-oxides precursor (BO) and conventional ceramic mixed-oxide methods (MO). Our experimental results show that the BO-type piezoceramics have better grain microstructure and better material properties [e.g., d(33)= 340 pc/N, k(t)= 0.52, Q(m)= 1250, temperature coefficient of change rate of resonant frequency (TCF) = 0.01%/degrees C, and temperature coefficient of change rate of clamped capacitance (TCC) = 0.18%/degrees C]. We construct 1-MHz transducers using our BO and MO types of piezoceramics and examine their dynamic resonant characteristics as the transducers are driven by a power driver with open-loop control. Results show that the transducers with the BO-type piezoceramics have better dynamic characteristics, such as time stability (e.g., aging rate of resonant frequency at thickness mode = 0.26%/decade cycle, and aging rate of clamped capacitance = 0.55%/decade cycle) and temperature stability as BO-type piezoceramics. Furthermore, we observe that the clamped capacitance variation has more influence on the transducer dynamic characteristics than the resonant frequency variation, and we verify the observation from the partial derivative ratio of the transfer function derived by a simulated ultrasonic equivalent circuit system. It is concluded that the BO-type piezoceramics are better candidates than the MO-type samples for obtaining optimum dynamic resonant characteristics in ultrasonic therapeutic transducers.


Description and comparison of excretory urography performed during radiography and computed tomography for evaluation of the urinary system in healthy New Zealand White rabbits (Oryctolagus cuniculus).
OBJECTIVE To evaluate the usefulness of excretory urography performed during radiography (REU) and CT (CTEU) in healthy rabbits, determine timings of urogram phases, and compare sensitivities of REU and CTEU for detection of these phases. ANIMALS 13 New Zealand White rabbits (Oryctolagus cuniculus). PROCEDURES Rabbits were screened for signs of systemic and urinary tract disease. An REU examination of each was performed, followed ≥ 5 days later by a CTEU examination. Contrast images from each modality were evaluated for quality of opacification and intervals between initiation of contrast medium administration and detection of various urogram phases. RESULTS Excretory urograms of excellent diagnostic quality were achieved with both imaging modalities. For all rabbits, the nephrographic phase of the urogram appeared in the first postcontrast REU image (obtained between 34 and 40 seconds after initiation of contrast medium administration) and at a median interval of 20 seconds in CTEU images. The pyelographic phase began at a median interval of 1.63 minutes with both imaging modalities. Contrast medium was visible within the urinary bladder at a median interval of 2.20 minutes. Median interval to the point at which the nephrogram and pyelogram were no longer visible in REU images was 8 hours and 2.67 hours, respectively. The CTEU technique was better than the REU technique for evaluating renal parenchyma. CONCLUSIONS AND CLINICAL RELEVANCE Findings suggested that REU and, particularly, CTEU may be valuable tools for the diagnosis of renal and urinary tract disease in rabbits; however, additional evaluation in diseased rabbits is required.


GSTT1 copy number gain is a poor predictive marker for escalated-dose imatinib treatment in chronic myeloid leukemia: genetic predictive marker found using array comparative genomic hybridization.
In a study population of 45 patients who were previously enrolled in an imatinib dose escalation trial, genome-wide screening for regions of genetic gains and losses was performed using array comparative genomic hybridization (aCGH). Early molecular response (EMR), defined as >50% reduction in the ratio of BCR-ABL1 to ABL1 within 6 months after dose escalation, was a major endpoint for analysis. After aCGH analysis, copy number change of four genes was investigated in 52 patients as a validation. Copy number gain in 16p11.2 was more frequently observed in patients with EMR than in patients who failed to achieve EMR (P = 0.034). A tendency for increased copy number in 22q11.23 in patients without EMR and for decreased copy number in 17q12 in patients with EMR was observed (P = 0.072 and P = 0.070, respectively). For GSTT1, in 22q11.23, copy number gain was observed in patients without EMR (P = 0.035). GSTT1 copy number gain was related to short time to treatment failure (TTFx) in patients without BCR-ABL1 mutations (P = 0.007). In multivariate analysis, GSTT1 copy number gain was an independent predictive factor for short TTFx (P = 0.020). We conclude that chromosome regions 16p11.2, 22q11.23, and 17q12 are potential locations related to response in imatinib dose escalation therapy for CML. GSTT1 copy number gain is a genetic change affecting outcome in this setting.


Subsidence Rates After Lateral Lumbar Interbody Fusion: A Systematic Review.
The evidence regarding the consequences of subsidence with lateral lumbar interbody fusion (LLIF) has been sparse. The objective of this study is to calculate the incidence of subsidence and reoperation for subsidence after LLIF. A secondary outcome examined the quantitative degree of subsidence by calculating the percent change in the height of the intervertebral space secondary to interbody subsidence at various postoperative follow-up times. Following the MOOSE (Meta-analysis [and Systematic Review] Of Observational Studies in Epidemiology) guidelines, a systematic review searched for all cohort studies that focused on subsidence rates after LLIF, including extreme lateral interbody fusions (XLIFs) and direct lateral interbody fusion. Neoplastic, infectious, and/or metabolic indications for LLIF were similarly excluded because these diseases may compromise bone quality and, thus, confound the rate of cage subsidence. Corpectomies were removed from the systematic review because 1) indications for removal of vertebral body typically reflect those excluded diseases and 2) subsidence refers to a different biomechanical process. This systematic review identified a subsidence incidence with LLIF of 10.3% (N = 141/1362 patients in 14 articles) and reoperation rate for subsidence of 2.7% (N = 41/1470 patients in 16 articles). In the secondary outcome measure, the disc height decreased from 5.6% after 3 months, 6.0% after 6 months, and 10.2% after 12 months, to 8.9% after 24 months (P < 0.001). Subsidence after LLIF carries a nonnegligible risk that may be incorporated in surgical consent discussions in selected patients.


Liver FOXP3 and PD1/PDL1 Expression is Down-Regulated in Chronic HBV Hepatitis on Maintained Remission Related to the Degree of Inflammation.
T cell expression of PD1 and inhibition of T effector cells by Foxp3(+)-T regulatory cells are among the most powerful mechanisms for achieving a balanced immune response. Our aim was to investigate, how liver FOXP3 and PD1/PDL1 expression is regulated in chronic HBV hepatitis (CHB) on maintained long-term remission in comparison with active disease, and whether they are correlated to the expression of pro- and anti-inflammatory cytokines and apoptosis mediators, along with the degree of histological inflammation and markers of T cell effector restoration. Fifty-three HBeAg-negative CHB patients with both active (30) and completely remitted disease on long-term antiviral treatment (23) and four controls (submitted to liver biopsy due to a mild increase of aminotransferases but without liver necroinflammatory and architecture changes) were enrolled in the study. Liver mRNA levels of immunoregulatory genes (FOXP3, IL10, TGFB1, and those of PD1/PDL1/PDL2 pathway), major apoptosis mediators (FAS, FASL, TNFA, TRAIL), cytokines of effector T cell restoration (IL2, IFNG), and those of IL1B, CD4, and CD8, were evaluated by quantitative real-time reverse-transcriptase PCR and were correlated with each other, along with the intensity of liver inflammation and fibrosis staging. The expression and localization of FOXP3, PD1, PDL1, CD4, and CD8 were also assessed by immunohistochemistry. The expression of FOXP3, IL10, TGFB1, PD1, PDL1, FASL, and CD8 was significantly down-regulated in the remission state. In contrast, liver expression of IL2 and IFNG, along with CD4, IL1B, TNFA, and FAS did not change significantly. Moreover, FOXP3, PD1, PDL1, and CD8 transcripts were positively correlated to the intensity of liver inflammation. Our data indicate that in the CHB disease model, the immunosuppressive liver environment is down-regulated in the maintained on-treatment long-term remission state and correlates with the intensity of liver inflammation, but not liver T cell restoration.


High-throughput screening with nucleosome substrate identifies small-molecule inhibitors of the human histone lysine methyltransferase NSD2.
The histone lysine methyltransferase nuclear receptor-binding SET domain protein 2 (NSD2, also known as WHSC1/MMSET) is an epigenetic modifier and is thought to play a driving role in oncogenesis. Both NSD2 overexpression and point mutations that increase its catalytic activity are associated with several human cancers. Although NSD2 is an attractive therapeutic target, no potent, selective, and bioactive small molecule inhibitors of NSD2 have been reported to date, possibly due to the challenges of developing high-throughput assays for NSD2. Here, to establish a platform for the discovery and development of selective NSD2 inhibitors, we optimized and implemented multiple assays. We performed quantitative high-throughput screening with full-length WT NSD2 and a nucleosome substrate against a diverse collection of bioactive small molecules comprising 16,251 compounds. We further interrogated 174 inhibitory compounds identified in the primary screen with orthogonal and counter assays and with activity assays based on the clinically relevant NSD2 variants E1099K and T1150A. We selected five confirmed inhibitors for follow-up, which included a radiolabeled validation assay, surface plasmon resonance studies, methyltransferase profiling, and histone methylation in cells. We found that all five NSD2 inhibitors bind the catalytic SET domain and one exhibited apparent activity in cells, validating the workflow and providing a template for identifying selective NSD2 inhibitors. In summary, we have established a robust discovery pipeline for identifying potent NSD2 inhibitors from small-molecule libraries.


The surgical treatment of traumatic brachial plexus lesions.
The vast majority of severe traumatic lesions of the brachial plexus are due to road accidents. Surgical intervention is indicated in cases of persistent motor and sensory loss. The operation should be performed one to three months after the traumatic event. Indications and results of neurolysis, autologous nerve grafts, and neurotization are reported. Neurolysis was employed 43 times on 36 patients primarily to relieve pain rather than to restore function. Autologous nerve grafting was employed in 100 patients and can give good results when the lesion involves the three upper roots. Neurotization using Bell's nerve was used in 46 patients as a source of axons to re-establish the nervous pathways of the destroyed suprascapular or musculocutaneous nerves.


Primary complex of paracoccidioidomycosis and hypereosinophilia.
Primary infection with Paracoccidioides brasiliensis has rarely been observed. A 28-year-old male patient presented with a three-month history of fever, respiratory symptoms and malaise. Chest X-rays revealed bilateral apical infiltrates, right pleuritis and hilar lymphadenomegaly. The patient presented with leukocytosis, severe eosinophilia and increasing titers of anti-P. brasiliensis antibodies in serum. To our knowledge, this is the first report of the primary pulmonary lymph node complex of paracoccidioidomycosis accompanied by hypereosinophilia and affecting a previously healthy adult.


Neuronal expression and subcellular localization of cholesterol 24-hydroxylase in the mouse brain.
Cholesterol 24-hydroxylase is a cytochrome P450 (CYP46A1) that is selectively expressed in the brain and is responsible for the majority of cholesterol turnover in the central nervous system. Mice deficient in 24-hydroxylase exhibit impaired learning and defective hippocampal long-term potentiation, suggesting that the metabolism of cholesterol by this enzyme is required for learning and memory formation. To determine where in the neuron cholesterol turnover was taking place, monoclonal antibodies directed against 24-hydroxylase were generated by immunization of mice with recombinant protein and used to detect the enzyme in brain homogenates, cultured neurons, and histological sections. 24-Hydroxylase was localized to the endoplasmic reticulum and was distributed throughout the cell bodies and dendrites of multiple types of neurons; the enzyme was not detected in axon terminals or in the cells of 24-hydroxylase knockout mice. 24-Hydroxylase was highly expressed in pyramidal neurons of the hippocampus and cortex, in Purkinje cells of the cerebellum, and in hippocampal and cerebellar interneurons. Within the retina, 24-hydroxylase was detected in ganglion cells and some but not all cells of the inner nuclear layer. These findings reveal the microsomal localization of 24-hydroxylase and provide subcellular insight into cholesterol turnover in the brain.


Neurological consequences of diabetic ketoacidosis at initial presentation of type 1 diabetes in a prospective cohort study of children.
To investigate the impact of new-onset diabetic ketoacidosis (DKA) during childhood on brain morphology and function. Patients aged 6-18 years with and without DKA at diagnosis were studied at four time points: <48 h, 5 days, 28 days, and 6 months postdiagnosis. Patients underwent magnetic resonance imaging (MRI) and spectroscopy with cognitive assessment at each time point. Relationships between clinical characteristics at presentation and MRI and neurologic outcomes were examined using multiple linear regression, repeated-measures, and ANCOVA analyses. Thirty-six DKA and 59 non-DKA patients were recruited between 2004 and 2009. With DKA, cerebral white matter showed the greatest alterations with increased total white matter volume and higher mean diffusivity in the frontal, temporal, and parietal white matter. Total white matter volume decreased over the first 6 months. For gray matter in DKA patients, total volume was lower at baseline and increased over 6 months. Lower levels of N-acetylaspartate were noted at baseline in the frontal gray matter and basal ganglia. Mental state scores were lower at baseline and at 5 days. Of note, although changes in total and regional brain volumes over the first 5 days resolved, they were associated with poorer delayed memory recall and poorer sustained and divided attention at 6 months. Age at time of presentation and pH level were predictors of neuroimaging and functional outcomes. DKA at type 1 diabetes diagnosis results in morphologic and functional brain changes. These changes are associated with adverse neurocognitive outcomes in the medium term.


Daily eating frequency and cardiometabolic risk factors in young Australian adults: cross-sectional analyses.
Eating frequency may be important in the development of overweight and obesity and other cardiometabolic risk factors; however, the evidence is inconsistent. The aim of the present study was to examine the associations between the number of eating occasions and cardiometabolic risk factors in a national population-based sample of young adults. A cohort of 1273 men and 1502 women, aged 26-36 years, completed a meal pattern chart to record when they had eaten during the previous day (in hourly intervals). The total number of eating occasions was calculated. Diet quality was assessed, waist circumference was measured and a fasting blood sample was taken. Dietary intake was compared with the Australian Guide to Healthy Eating. The associations between the number of eating occasions and cardiometabolic risk factors were calculated using linear regression. Analyses were adjusted for age, education and physical activity. Most men ate three to five times per d and most women ate four to six times. The proportion of participants meeting dietary recommendations increased with the number of eating occasions. For men, an additional eating occasion was associated with reductions in mean values for waist circumference (-0·75 cm), fasting glucose (-0·02 mmol/l), fasting insulin (-0·34 mU/l; 2·04 pmol/l), TAG (-0·03 mmol/l), total cholesterol (-0·08 mmol/l) and LDL-cholesterol (-0·06 mmol/l). Adjustment for waist circumference attenuated the results. Significant trends were not observed for women. In conclusion, a higher number of eating occasions were associated with reduced cardiometabolic risk factors in men. Many associations were mediated by waist circumference.


Difficulties with invasive risk stratification performed under anesthesia in pediatric Wolff-Parkinson-White Syndrome.
Children with Wolff-Parkinson-White Syndrome (WPW) are at risk for sudden death. The gold standard for risk stratification in this population is the shortest pre-excited RR interval during atrial fibrillation (SPERRI). The purpose of this study was to determine how closely measurements made in the electrophysiology laboratory in patients with WPW compared to SPERRI obtained during an episode of clinical pre-excited atrial fibrillation (Clinical-SPERRI). This was a subgroup analysis of a multicenter study of children with WPW. Subjects in our study (N = 49) were included if they had Clinical-SPERRI measured in addition to 1 or more of 3 surrogate measurements: SPERRI obtained during electrophysiological study (EP-SPERRI), accessory pathway effective refractory period (APERP), or shortest pre-excited paced cycle length with 1:1 conduction (SPPCL). Seventy percent of electrophysiological measurements were made with patients under general anesthesia. Clinical-SPERRI moderately correlated with EP-SPERRI (r = 0.495; P = .012). However, 24% of our patients with Clinical-SPERRI ≤250 ms would have been misclassified as having a low-risk pathway based on EP-SPERRI >250 ms. Clinical-SPERRI did not correlate with APERP or SPPCL (r < 0.3; P >.1). Mean EP-SPERRI, APERP, and SPPCL all were greater than Clinical-SPERRI. Electrophysiology laboratory measurements of pathway characteristics made with patients under general anesthesia do not correlate well with Clinical-SPERRI. Of APERP, SPPCL, and EP-SPERRI, only EP-SPERRI had moderate correlation with Clinical-SPERRI. This study questions the predictive ability of invasive risk stratification with patients under general anesthesia, given that 24% of patients with high-risk Clinical-SPERRI (≤250 ms) had EP-SPERRI that may be considered low risk (>250 ms).


Chemical aerosol flow synthesis of semiconductor nanoparticles.
Nanometer-sized semiconductor particles (quantum dots) have been the subject of intense research during the past decade owing to their novel electronic, catalytic, and optical properties. Fundamental properties of these nanoparticles (1-20 nm diameter) can be systematically changed simply by controlling the size of the crystals while holding their chemical composition constant. We describe here a new methodology for the continuous production of fluorescent CdS, CdSe, and CdTe nanoparticles using ultrasonically generated aerosols of high boiling point solvents. Each submicron droplet serves as a separate nanoscale chemical reactor, with reactions proceeding as the liquid droplets (which hold both reactants and surface stabilizers) are heated in a gas stream. The method is inexpensive, scalable, and allows for the synthesis of high quality nanocrystals. This chemical aerosol flow synthesis (CAFS) can be extended to the synthesis of nanostructured metals, oxides, and other materials.


Phase and light shift determination in an optically pumped cesium beam frequency standard.
The determination of the phase shift and the light shift in our optically pumped Cs beam is reported here. We show that an error on the phase difference value determined by our fit method, without beam reversal, can be induced if the light shift is not taken into account. If the experimental data are corrected for the light shift, the results of the fit method are in very good agreement with the beam reversal results. This allows us to reduce the related uncertainty in the accuracy budget of our standard. The resulting overall accuracy of the standard is now estimated to be 6.3x10(-15).


Asymmetric reproductive barriers and mosaic reproductive isolation: insights from Misty lake-stream stickleback.
Ecological speciation seems to occur readily but is clearly not ubiquitous - and the relative contributions of different reproductive barriers remain unclear in most systems. We here investigate the potential importance of selection against migrants in lake/stream stickleback (Gasterosteus aculeatus) from the Misty Lake system, Canada. This system is of particular interest because one population contrast (Lake vs. Outlet stream) shows very low genetic and morphological divergence, whereas another population contrast (Lake vs. Inlet stream) shows dramatic genetic and morphological divergence apparently without strong and symmetric reproductive barriers. To test whether selection against migrants might solve this "conundrum of missing reproductive isolation", we performed a fully factorial reciprocal transplant experiment using 225 individually marked stickleback collected from the wild. Relative fitness of the different ecotypes (Lake, Inlet, and Outlet) was assessed based on survival and mass change in experimental enclosures. We found that Inlet fish performed poorly in the lake (selection against migrants in that direction), whereas Lake fish outperformed Inlet fish in all environments (no selection against migrants in the opposite direction). As predicted from their phenotypic and genetic similarity, Outlet and Lake fish performed similarly in all environments. These results suggest that selection against migrants is asymmetric and, together with previous work, indicates that multiple reproductive barriers contribute to reproductive isolation. Similar mosaic patterns of reproductive isolation are likely in other natural systems.


Cytotoxic activity of C-geranyl compounds from Paulownia tomentosa fruits.
The newly discovered 5,7-dihydroxy-6-geranylchromone ( 1) was isolated from PAULOWNIA TOMENTOSA fruit and subsequently characterized. The structure of the isolated compound was elucidated on the basis of extensive NMR experiments including HMQC, HMBC, COSY, and NOESY, as well as HR-MS, IR, and UV. The cytotoxicity of 1 was evaluated using a plant cell model represented by tobacco BY-2 cells. The other phytoconstituents ( 2 - 8) previously isolated from P. TOMENTOSA were similarly evaluated together with the known flavanones 10 and 11. The cytotoxicity (human erythro-leukaemia cell line K562) and activity on erythroid differentiation of compounds 2 - 9 and 12 and 13 have also been evaluated. Acteoside ( 2) was determined to be the most toxic of the compounds tested on BY-2 cells, diplacone ( 6) on the K562 cell line. Some aspects of the relationship between the flavanone skeleton substitution and the metabolic activation necessary for a toxic effect are discussed.


Regulations of astrocytic functions by endothelins: roles in the pathophysiological responses of damaged brains.
The receptors for endothelins (ETs) are classified into the ET(A) and ET(B) types. ET(B) receptors are highly expressed in astrocytes, but pharmacological usages of this receptor are not clarified. In this article, recent studies on the pathophysiological roles of astrocytic ET(B) receptors in the brain are reviewed. The administration of ET(B) agonists and antagonists in nerve injury models showed that several astrocytic functions are regulated by ET(B) receptors. The activation of ET(B) receptors causes morphological alterations and proliferation of cultured astrocytes. Astrocytes produce various bio-active substances that can affect damage to nerve tissues. ETs stimulate the production of neurotrophic factors by astrocytes. This action improves impaired brain functions. On the other hand, the production of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF), which induce brain edema, also are stimulated by ETs. These findings indicate that astrocytic functions are effectively regulated by modulations of ET(B) receptors. In brain insults and neurodegenerative diseases, these functions of astrocytes affect the protection and repair of damaged nerve tissues. Thus, astrocytic ET(B) receptors could be a target for novel types of neuroprotective drugs.


[Cumulative effect of alcohol (ethanol) on the activity of the cerebellar Purkinje cells of the cat].
The activity of the identified cerebellar Purkinje cells after three successive intravenous injections of 2 ml/kg of 30% alcohol (ethanol) solution was studied in anaesthetized cats. After the first injection a sharp increase in the discharge frequency of Purkinje cells activated by mossy fibre afferent input was accompanied by a decrease in the discharge frequency of the Purkinje cells activated by climbing fiber input. Each successive injection of the same dose of alcohol produced a cumulative effect on the activity of the cerebellar Purkinje cells.


Neural correlates of heart rate variability in PTSD during sub- and supraliminal processing of trauma-related cues.
Posttraumatic stress disorder (PTSD) is characterized by dysregulated arousal and altered cardiac autonomic response as evidenced by decreased high-frequency heart rate variability (HF-HRV), an indirect measure of parasympathetic modulation of the heart. Indeed, subtle threatening cues can cause autonomic dysregulation, even without explicit awareness of the triggering stimulus. Accordingly, examining the neural underpinnings associated with HF-HRV during both sub- and supraliminal exposure to trauma-related cues is critical to an enhanced understanding of autonomic nervous system dysfunction in PTSD. We compared neural activity in brain regions associated with HF-HRV in PTSD (n = 18) and healthy controls (n = 18) during exposure to sub- and supraliminal processing of personalized trauma-related words. As compared to controls, PTSD exhibited decreased HF-HRV reactivity in response to sub- and supraliminal cues. Notably, during subliminal processing of trauma-related versus neutral words, as compared to controls, PTSD showed decreased neural response associated with HF-HRV within the left dorsal anterior insula. By contrast, during supraliminal processing of trauma-related versus neutral words, decreased neural activity associated with HF-HRV within the posterior insula/superior temporal cortex, and increased neural activity associated with HF-HRV within the left centromedial amygdala was observed in PTSD as compared to controls. Impaired parasympathetic modulation of autonomic arousal in PTSD appears related to altered activation of cortical and subcortical regions involved in the central autonomic network. Interestingly, both sub- and supraliminal trauma-related cues appear to elicit dysregulated arousal and may contribute to the maintenance of hyperarousal in PTSD. Hum Brain Mapp 38:4898-4907, 2017. © 2017 Wiley Periodicals, Inc.


siRNA screen identifies QPCT as a druggable target for Huntington's disease.
Huntington's disease (HD) is a currently incurable neurodegenerative condition caused by an abnormally expanded polyglutamine tract in huntingtin (HTT). We identified new modifiers of mutant HTT toxicity by performing a large-scale 'druggable genome' siRNA screen in human cultured cells, followed by hit validation in Drosophila. We focused on glutaminyl cyclase (QPCT), which had one of the strongest effects on mutant HTT-induced toxicity and aggregation in the cell-based siRNA screen and also rescued these phenotypes in Drosophila. We found that QPCT inhibition induced the levels of the molecular chaperone αB-crystallin and reduced the aggregation of diverse proteins. We generated new QPCT inhibitors using in silico methods followed by in vitro screening, which rescued the HD-related phenotypes in cell, Drosophila and zebrafish HD models. Our data reveal a new HD druggable target affecting mutant HTT aggregation and provide proof of principle for a discovery pipeline from druggable genome screen to drug development.


PathOS: a decision support system for reporting high throughput sequencing of cancers in clinical diagnostic laboratories.
The increasing affordability of DNA sequencing has allowed it to be widely deployed in pathology laboratories. However, this has exposed many issues with the analysis and reporting of variants for clinical diagnostic use. Implementing a high-throughput sequencing (NGS) clinical reporting system requires a diverse combination of capabilities, statistical methods to identify variants, global variant databases, a validated bioinformatics pipeline, an auditable laboratory workflow, reproducible clinical assays and quality control monitoring throughout. These capabilities must be packaged in software that integrates the disparate components into a useable system. To meet these needs, we developed a web-based application, PathOS, which takes variant data from a patient sample through to a clinical report. PathOS has been used operationally in the Peter MacCallum Cancer Centre for two years for the analysis, curation and reporting of genetic tests for cancer patients, as well as the curation of large-scale research studies. PathOS has also been deployed in cloud environments allowing multiple institutions to use separate, secure and customisable instances of the system. Increasingly, the bottleneck of variant curation is limiting the adoption of clinical sequencing for molecular diagnostics. PathOS is focused on providing clinical variant curators and pathology laboratories with a decision support system needed for personalised medicine. While the genesis of PathOS has been within cancer molecular diagnostics, the system is applicable to NGS clinical reporting generally. The widespread availability of genomic sequencers has highlighted the limited availability of software to support clinical decision-making in molecular pathology. PathOS is a system that has been developed and refined in a hospital laboratory context to meet the needs of clinical diagnostics. The software is available as a set of Docker images and source code at https://github.com/PapenfussLab/PathOS .


Evolution in techniques: lip augmentation.
Lip augmentation is an increasingly popular procedure. With increasing demand from patients comes an increasing challenge to surgeons to evolve techniques that are suited to the particular concerns, desires, and anatomy of each patient. In the past, options were limited to only a few filler substances and relatively few surgical options. Today new filler products continue to rapidly evolve and become available to surgeons and are often directly marketed to patients via various media formats. Tissue grafts and autologous fat grafting are also giving surgeons options for lip enhancement that were limited in the past. Options for permanent lip augmentation with newly advanced alloplastic implants are now available to help patients achieve a more natural look and feel. The challenge for surgeons of any level of experience is to weigh the advantages and disadvantages of each newly emerging technique and incorporate these into his or her armamentarium. I will outline the evolution of my lip augmentation techniques, emphasizing the use of Advanta for permanent lip augmentation, and the rationale behind this evolution.


Comparison of vaginoscopic hysteroscopy and traditional hysteroscopy: A systematic review and meta-analysis.
Outpatient hysteroscopy is a safe, feasible, and optimum procedure for the diagnosis and management of intrauterine pathologies. To determine the best approach of outpatient hysteroscopy (vaginoscopic vs traditional) in terms of pain, duration of procedure, feasibility, safety, and acceptability. PubMed, Embase, Google Scholar, and Scopus were searched from January 2000 to October 2021. No filters or restrictions were applied. Randomized controlled trials comparing vaginoscopic hysteroscopy with traditional hysteroscopy in an outpatient setting. Two authors independently performed a comprehensive literature search and collected and extracted data. The summary effect estimate was determined using both fixed effects and random-effects models. Seven studies with 2723 patients (vaginoscopic [n = 1378] and traditional hysteroscopy [n = 1345]) were included. Vaginoscopic hysteroscopy was associated with a significant reduction in intraprocedural pain (standardized mean difference, -0.05 [95% confidence interval (CI), -0.33 to -0.23], I2 = 0%), procedural time (standardized mean difference, -0.45 [95% CI, -0.76 to -0.14], I2 = 82%), and fewer side effects (relative risk, 0.37 [95% CI, 0.15-0.91], I2 = 0%). The procedure failure rate was similar in both approaches (relative risk, 0.97 [95% CI, 0.71-1.32], I2 = 43%). Complications were mostly documented with traditional hysteroscopy. Vaginoscopic hysteroscopy reduces the pain and duration compared with traditional hysteroscopy.


Effect of BI-1 on insulin resistance through regulation of CYP2E1.
Diet-induced obesity is a major contributing factor to the progression of hepatic insulin resistance. Increased free fatty acids in liver enhances endoplasmic reticulum (ER) stress and production of reactive oxygen species (ROS), both are directly responsible for dysregulation of hepatic insulin signaling. BI-1, a recently studied ER stress regulator, was examined to investigate its association with ER stress and ROS in insulin resistance models. To induce obesity and insulin resistance, BI-1 wild type and BI-1 knock-out mice were fed a high-fat diet for 8 weeks. The BI-1 knock-out mice had hyperglycemia, was associated with impaired glucose and insulin tolerance under high-fat diet conditions. Increased activity of NADPH-dependent CYP reductase-associated cytochrome p450 2E1 (CYP2E1) and exacerbation of ER stress in the livers of BI-1 knock-out mice was also observed. Conversely, stable expression of BI-1 in HepG2 hepatocytes was shown to reduce palmitate-induced ER stress and CYP2E1-dependent ROS production, resulting in the preservation of intact insulin signaling. Stable expression of CYP2E1 led to increased ROS production and dysregulation of insulin signaling in hepatic cells, mimicking palmitate-mediated hepatic insulin resistance. We propose that BI-1 protects against obesity-induced hepatic insulin resistance by regulating CYP2E1 activity and ROS production.


Evaluation of the Char Formation During the Hydrothermal Treatment of Wooden Balls.
With wooden balls, a visualization of the hydrothermal carbonization to show the progress of the conversion to char is presented. In the present study, the balls represent the particles of biomass to investigate the differences in conversion outside and inside of biomass particles, during hydrothermal carbonization. A special focus is on hydrochar and pyrochar formation. The wooden balls are treated in subcritical water at 220 °C for holding times between 0 and 960 min. Even after 960 min, hydrolysis of the original biomass is incomplete as cellulose and hemicellulose are linked by lignin, inhibiting the reaction with water. Moreover, two different pathways of char production can be observed. Inside of the wooden ball pyrochar is formed as any water got that deep in, on the surface hydrochar is fixed, originated from the surrounding liquid. On the ground of the HTC reactor, a thin, brittle precipitate of likely hydrochar or humins can be found either from the precipitation of loosely attached compounds on the surface of the biomass or direct precipitation from the liquid.


Changes in nutrient concentrations of leaves and roots in response to global change factors.
Global change impacts on biogeochemical cycles have been widely studied, but our understanding of whether the responses of plant elemental composition to global change drivers differ between above- and belowground plant organs remains incomplete. We conducted a meta-analysis of 201 reports including 1,687 observations of studies that have analyzed simultaneously N and P concentrations changes in leaves and roots in the same plants in response to drought, elevated [CO2 ], and N and P fertilization around the world, and contrasted the results within those obtained with a general database (838 reports and 14,772 observations) that analyzed the changes in N and P concentrations in leaves and/or roots of plants submitted to the commented global change drivers. At global level, elevated [CO2 ] decreased N concentrations in leaves and roots and decreased N:P ratio in roots but no in leaves, but was not related to P concentration changes. However, the response differed among vegetation types. In temperate forests, elevated [CO2 ] was related with lower N concentrations in leaves but not in roots, whereas in crops, the contrary patterns were observed. Elevated [CO2 ] decreased N concentrations in leaves and roots in tundra plants, whereas not clear relationships were observed in temperate grasslands. However, when elevated [CO2 ] and N fertilization coincided, leaves had lower N concentrations, whereas root had higher N concentrations suggesting that more nutrients will be allocated to roots to improve uptake of the soil resources not directly provided by the global change drivers. N fertilization and drought increased foliar and root N concentrations while the effects on P concentrations were less clear. The changes in N and P allocation to leaves and root, especially those occurring in opposite direction between them have the capacity to differentially affect above- and belowground ecosystem functions, such as litter mineralization and above- and belowground food webs.


Microsatellite instability is uncommon in young patients with renal cell carcinoma.
Hereditary non-polyposis colorectal cancer (HNPCC) is an autosomal, dominantly inherited syndrome which predisposes to the development of colorectal cancer and, to a lesser extent, other extracolonic neoplasms. Widespread microsatellite instability is a feature of nearly all tumors from patients with hereditary non-polyposis colorectal cancer. It was recently found that most patients under age 35 with colorectal cancer also have widespread microsatellite instability in their tumors and that many of these patients have the germline mutations of mismatch repair genes that characterize hereditary non-polyposis colorectal cancer. Renal cell carcinoma has occasionally been reported in patients with hereditary non-polyposis colorectal cancer, and some renal cell carcinomas have been shown to have microsatellite instability at multiple loci. For this reason, we sought to address the possibility that some young patients with renal cell carcinoma may have hereditary non-polyposis colorectal cancer by screening their tumors for microsatellite instability. Thirty-two patients under the age of 45 with renal cell carcinoma were examined for the presence of microsatellite instability in their tumor tissue. Microsatellite instability was not observed in any of the tumors. The absence of microsatellite instability in renal cell carcinomas from young patients suggests that these patients are unlikely to have hereditary non-polyposis colorectal cancer and would generally not benefit from genetic screening for this syndrome.


Diagnostic accuracy of fluorescence microlymphography for detecting limb lymphedema.
Fluorescence microlymphography (FML) is a minimally invasive technique for visualization of the cutaneous lymphatic network. The aim of the study was to assess the accuracy and safety of FML in patients with unilateral lymphedema. This was a cross sectional study. Patients with unilateral leg swelling were assessed and compared with the unaffected contralateral limb. FML was performed in all index legs and the contralateral leg by injecting 0.1 mL of fluorescein isothiocyanate (FITC)-labeled dextran intradermally in both limbs at the same level. The most prominent swelling of the affected limb was the anatomical reference. The spread of the dye in the lymphatic capillaries of the skin was measured in all dimensions by epiluminator intravital microscopy and the maximum dye spread value 10 min after injection was used for statistical analysis. The contralateral leg served as control. Test accuracy and receiver operator characteristic (ROC) analysis was performed to assess threshold values that best predict lymphedema. Between March 2008 and February 2014 seventy patients with unilateral chronic leg swelling were clinically diagnosed with lymphedema. The median age was 45 (IQR 27-56) years. Of those, 46 (65.7%) were female and 71.4% had primary and 28.6% secondary lymphedema. Sensitivity, specificity, positive and negative likelihood ratio, and positive and negative predictive value were 94.3%, 78.6%, 4.40, 0.07, 81.5%, and 93.2% for the 12 mm cut off level and 91.4%, 85.7%, 6.40, 0.10, 86.5%, and 90.9% for the 14 mm cut off level, respectively. The area under the ROC curve was 0.89 (95% CI: 0.83-0.95). No major adverse events were observed. FML is an almost atraumatic and safe technique for detecting lymphedema in patients with leg swelling. In this series the greatest accuracy was observed at a cut off level of ≥14 mm maximum spread.


Four-year epidemiological surveillance of the Spanish Registry of Contact Dermatitis and Cutaneous Allergy (REIDAC): Current situation and trends.
The epidemiological surveillance of contact dermatitis is one of the objectives of the Spanish Registry of Contact Dermatitis and Cutaneous Allergy (REIDAC). Knowing whether the prevalence of positive tests to the different allergens changes over time is important for this monitoring process. To describe the various temporary trends in allergen positivity in the GEIDAC standard series from 2018 through December 31, 2022. This was a multicenter, observational trial of consecutive patients analyzed via patch tests as part of the study of possible allergic contact dermatitises collected prospectively within the REIDAC. The data was analyzed using 2 statistical tests: one homogeneity test (to describe the changes seen over time) and one trend test (to see whether the changes described followed a linear trend). A total of 11 327 patients were included in the study. Overall, the allergens associated with a highest sensitization were nickel sulfate, methylisothiazolinone, cobalt chloride, methylchloroisothiazolinone/methylisothiazolinone, and fragrance mix I. A statistically significant decrease was found in the percentage of methylisothiazolinone positive tests across the study years with an orderly trend. Although various changes were seen in the sensitizations trends to several allergens of the standard testing, it became obvious that a high sensitization to nickel, methylchloroisothiazolinone/methylisothiazolinone and fragrances mix I remained. Only a significant downward trend was seen for methylisothiazolinone.


Activation of invariant NKT cells ameliorates experimental ocular autoimmunity by a mechanism involving innate IFN-gamma production and dampening of the adaptive Th1 and Th17 responses.
Invariant NKT cells (iNKT cells) have been reported to play a role not only in innate immunity but also to regulate several models of autoimmunity. Furthermore, iNKT cells are necessary for the generation of the prototypic eye-related immune regulatory phenomenon, anterior chamber associated immune deviation (ACAID). In this study, we explore the role of iNKT cells in regulation of autoimmunity to retina, using a model of experimental autoimmune uveitis (EAU) in mice immunized with a uveitogenic regimen of the retinal Ag, interphotoreceptor retinoid-binding protein. Natural strain-specific variation in iNKT number or induced genetic deficiencies in iNKT did not alter baseline susceptibility to EAU. However, iNKT function seemed to correlate with susceptibility and its pharmacological enhancement in vivo by treatment with iNKT TCR ligands at the time of uveitogenic immunization reproducibly ameliorated disease scores. Use of different iNKT TCR ligands revealed dependence on the elicited cytokine profile. Surprisingly, superior protection against EAU was achieved with alpha-C-GalCer, which induces a strong IFN-gamma but only a weak IL-4 production by iNKT cells, in contrast to the ligands alpha-GalCer (both IFN-gamma and IL-4) and OCH (primarily IL-4). The protective effect of alpha-C-GalCer was associated with a reduction of adaptive Ag-specific IFN-gamma and IL-17 production and was negated by systemic neutralization of IFN-gamma. These data suggest that pharmacological activation of iNKT cells protects from EAU at least in part by a mechanism involving innate production of IFN-gamma and a consequent dampening of the Th1 as well as the Th17 effector responses.


[Implementation of AUGE plan for chronic renal failure patients. The implementation of Universal Access and Explicit Guaranties Plan (AUGE), generates longer waiting lists among patients with chronic renal failure].
An opportune, efficient and low cost medical attention should be provided to all patients, without discrimination. When the need for medical attention outweighs the available resources, permanently increasing waiting lists are generated. The aim of the Universal Access and Explicit Guaranties Plan (AUGE) is to attend selected patients in periods predefined by the health authority. To describe the first six months of implementation of AUGE plan for chronic renal failure in a public hospital of Santiago. As a health care quality monitoring system, the lapse between the generation of a nephrological consultation and the specialist assessment, is registered periodically. These lapses were evaluated before and after the AUGE plan implementation, for eligible and non eligible patients for the plan. Results were modeled according queue theory. After three months of implementation, the AUGE plan prolongs waiting periods and increases the number of all patients seeking for nephrological attention. According to these results AUGE plan will be successful only if its present functioning is modified and receives more financial resources.


Nitro-oleic acid inhibits angiotensin II-induced hypertension.
Nitro-oleic acid (OA-NO(2)) is a bioactive, nitric-oxide derived fatty acid with physiologically relevant vasculoprotective properties in vivo. OA-NO(2) exerts cell signaling actions as a result of its strong electrophilic nature and mediates pleiotropic cell responses in the vasculature. The present study sought to investigate the protective role of OA-NO(2) in angiotensin (Ang) II-induced hypertension. We show that systemic administration of OA-NO(2) results in a sustained reduction of Ang II-induced hypertension in mice and exerts a significant blood pressure lowering effect on preexisting hypertension established by Ang II infusion. OA-NO(2) significantly inhibits Ang II contractile response as compared to oleic acid (OA) in mesenteric vessels. The improved vasoconstriction is specific for the Ang II type 1 receptor (AT(1)R)-mediated signaling because vascular contraction by other G-protein-coupled receptors is not altered in response to OA-NO(2) treatment. From the mechanistic viewpoint, OA-NO(2) lowers Ang II-induced hypertension independently of peroxisome proliferation-activated receptor (PPAR)gamma activation. Rather, OA-NO(2), but not OA, specifically binds to the AT(1)R, reduces heterotrimeric G-protein coupling, and inhibits IP(3) (inositol-1,4,5-trisphosphate) and calcium mobilization, without inhibiting Ang II binding to the receptor. These results demonstrate that OA-NO(2) diminishes the pressor response to Ang II and inhibits AT(1)R-dependent vasoconstriction, revealing OA-NO(2) as a novel antagonist of Ang II-induced hypertension.


A trans-ethnic genetic study of rheumatoid arthritis identified FCGR2A as a candidate common risk factor in Japanese and European populations.
Rheumatoid arthritis (RA) is a common systemic autoimmune disease and its onset and prognosis are controlled by genetic, immunological, and environmental factors. The HLA locus, particularly HLA-DRB1, is its strongest genetic risk determinant across ethnicities. Several other genes, including PTPN22 and PADI4, show modest association with RA. However, they cover only a part of its genetic components and their relative contribution is different between populations. To identify novel genetic determinants, we took a candidate gene approach in a trans-ethnic manner. After critical selection of 169 genes based on their immunological function, we performed SNP discovery of these genes by the resequencing of exons and surrounding areas using European and Japanese DNAs. We then generated a panel of 1,509 SNPs for case-control association study in both populations. The DerSimonian-Laird test for meta-analysis, using the combined results of the two populations, identified rs7551957 at the 5'-flanking region of the low-affinity Fc-gamma receptor IIa (FCGR2A) gene as the strongest candidate for the association (p = 8.6 × 10(-5), odds ratio = 1.58 with 95%CI 1.25-1.99). Suggestive signals were also obtained for three SNPs in the dihydropyrimidine dehydrogenase (DPYD) gene (rs6685859; p = 1.3 × 10(-4), rs7550959; p = 1.5 × 10(-4) and rs7531138; p = 1.7 × 10(-4)) and an intronic SNP, rs2269310, of the erythrocytic spectrin beta (SPTB) gene (p = 7.9 × 10(-4)).


The impact of child sexual abuse on the course of bipolar disorder: a systematic review.
The aim of this review was to elucidate the impact of child sexual abuse on all clinical phenomena that occur after the onset of bipolar disorder, including associated clinical features that are not part of the diagnostic criteria for the disorder. Five databases were searched and supplemented with a hand search of reference lists from retrieved papers. Study quality was assessed using a validated quality assessment tool. Blind assessments of study eligibility and quality were conducted by two independent researchers to reduce bias, minimize errors, and enhance the reliability of findings. Disagreements were resolved by consensus. Eighteen studies that included a total of 2996 adults and youths with bipolar disorder and met the minimum quality criteria necessary to ensure objectivity and not invalidate results were analyzed. Across studies, child sexual abuse was strongly (and perhaps directly) associated with posttraumatic stress disorder; whereas it was less strongly (and perhaps indirectly) related to suicide attempts, alcohol and/or drug abuse or dependence, psychotic symptoms, and an early age of illness onset. In regard to the association between child sexual abuse and other clinical variables concerning the course of bipolar disorder, evidence was scant or conflicting. Child sexual abuse is associated (either directly or indirectly) with some clinical phenomena that represent a more severe form of bipolar disorder. Although such a traumatic experience may directly affect the development of posttraumatic stress disorder, the effects of early sexual abuse on later suicidal behavior, substance abuse, and psychotic symptoms may operate through the mediating influences of certain psychopathological or neurobiological variables.


Introduction to egg pasteurization.
The science of microbiology is very young in comparison with the age of mankind. Related developments in the area of pasteurization have been made during the past 131 yr. This symposium reveals some recent advancements in pasteurization of shell eggs and egg products.


Klatskin tumour: radical and palliative treatment.
Primitive neoplasms of proximal extrahepatic bile ducts keep their peculiar morphological and spreading patterns, as firstly described by J. Klatskin. Diagnostic and therapeutic approach were modified in the last year, on the basis of technological progress in imaging and more aggressive surgical attitude. Authors reviewed their clinical experience from 1970 to 1995 concerning proximal extrahepatic bile ducts tumors management, mainly evaluating the evolution of diagnostic work-up and the role of resection. Preoperative work-up is now trimed to non invasive techniques, in order to evaluate the intra and extra biliary diffusion; PTC-PTBD performed preoperatively give a clear biliary map, and could be the first step of a palliative definitive treatment in case of non operable patients. Radical resection remains the gold standard of therapy, with the best long-term results. Palliation must be obtained by the easiest comfortable method for the patient (i.e. self-blocking percutaneously inserted endoprosthesis).


Multipolar Ewald methods, 2: applications using a quantum mechanical force field.
A fully quantum mechanical force field (QMFF) based on a modified "divide-and-conquer" (mDC) framework is applied to a series of molecular simulation applications, using a generalized Particle Mesh Ewald method extended to multipolar charge densities. Simulation results are presented for three example applications: liquid water, p-nitrophenylphosphate reactivity in solution, and crystalline N,N-dimethylglycine. Simulations of liquid water using a parametrized mDC model are compared to TIP3P and TIP4P/Ew water models and experiment. The mDC model is shown to be superior for cluster binding energies and generally comparable for bulk properties. Examination of the dissociative pathway for dephosphorylation of p-nitrophenylphosphate shows that the mDC method evaluated with the DFTB3/3OB and DFTB3/OPhyd semiempirical models bracket the experimental barrier, whereas DFTB2 and AM1/d-PhoT QM/MM simulations exhibit deficiencies in the barriers, the latter for which is related, in part, to the anomalous underestimation of the p-nitrophenylate leaving group pKa. Simulations of crystalline N,N-dimethylglycine are performed and the overall structure and atomic fluctuations are compared with the experiment and the general AMBER force field (GAFF). The QMFF, which was not parametrized for this application, was shown to be in better agreement with crystallographic data than GAFF. Our simulations highlight some of the application areas that may benefit from using new QMFFs, and they demonstrate progress toward the development of accurate QMFFs using the recently developed mDC framework.


[Investigation of West Nile virus in central nervous system infections of unknown etiology in Ankara, Turkey].
Arthropod-borne viral infections have recently gained considerable attention and importance as re-emerging infections in a global scale. West Nile Virus (WNV), a member of Flaviviridae, is an enveloped positive strand RNA virus that is usually transmitted to humans by the bite of Culicine mosquitoes. Although the majority of the human infections are asymptomatic, WNV may also cause febrile and neuro-invasive diseases. Seroprevalence data from Turkey indicate that WNV activity is present in Central Anatolia. In this study performed at Hacettepe University Hospital, paired serum and cerebrospinal fluid (CSF) samples from 87 adult patients with the preliminary diagnosis of aseptic meningitis/encephalitis of unknown etiology were evaluated retrospectively to identify WNV-related syndromes. Bacterial, fungal and mycobacterial cultures yielded negative results and Mycobacterium tuberculosis and Herpes simplex virus nucleic acid tests were also negative for the selected patients. Commercial enzyme-linked immunosorbent assay (ELISA)s and indirect immunofluorescence test (IIFT)s were employed for WNV IgM and IgG antibody detection (Anti-WNV Virus IgG/IgM ELISA, Anti-WNV Virus IgG/IgM IIFT; Euroimmun, Germany). Additional ELISA/IIFT assays were further performed for WNV antibody reactive samples to identify cross-reactions and/or infections with other flaviviruses and phleboviruses. All WNV antibody positive samples were also evaluated by a WNV real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. WNV IgM and IgG antibodies were detected in %9.2 (8/87) and 3.4% (3/87) of the serum samples, respectively. All IgG reactive samples were negative for IgM. All sera with WNV antibody reactivity (n = 11) and the corresponding CSF samples were negative for viral RNA via RT-PCR. In 5 of the 8 WNV IgM positive subjects, sandfly fever virus IgM antibodies were detected, which was also accompanied by Dengue virus IgM positivity in one sample. In another case, intrathecal antibody synthesis against measles virus was demonstrated. Two cases (2/87; 2.3%) with WNV IgM positivity as the only serologic marker were identified as probable WNV infections and clinical features were discussed. In conclusion, in order to fully understand the impact of WNV and/or other flavivirus infections in Turkey, epidemiology and ecological features of these agents need to established.


Lumbar drainage and delayed cerebral ischemia in aneurysmal subarachnoid hemorrhage: a systematic review.
Lumbar drainage for cerebrospinal fluid (CSF) diversion in aneurysmal subarachnoid hemorrhage (aSAH) has been reported to be beneficial in small series. There is no consensus regarding the optimal candidates for lumbar drainage, timing of drain placement, or amount and duration of CSF drainage. We performed a comprehensive review of the English literature reporting series of patients with aSAH undergoing CSF diversion with lumbar drains. Favorable clinical outcome was defined as modified Rankin Scale of 0-2 or Glasgow Outcome Scale as 4-5. A total of 8 studies reporting on 841 patients were included. Of these, 446 patients were treated with lumbar drains. Two studies were prospective and five studies had comparison groups. Most patients undergoing lumbar drainage were in good clinical grade on presentation (394/446, 88%) and the majority had substantial clot burden on head CT. Among the five studies with a comparison group, lumbar drainage was associated with lower rates of symptomatic vasospasm or delayed cerebral ischemia (20% vs. 45%, P<0.001) and higher rates of favorable outcome (79.4% vs. 60.4% P<0.001). The complication rate was 3.5%. Lumbar drainage in aSAH appears to be safe and associated with reduced rates of symptomatic vasospasm and improved clinical outcomes in patients in good clinical grade with thick clot burden, but the quality of most available studies is weak. The optimal duration and rate of CSF diversion remains uncertain.


Development of the gait outcomes assessment list for lower-limb differences (GOAL-LD) questionnaire: a child and parent reported outcome measure.
To develop a priority-based patient/parent reported outcome measure for children with lower-limb differences (LD) by adapting the Gait Outcomes Assessment List (GOAL) questionnaire. Guided by a conceptual framework of patient priorities, the GOAL questionnaire was iteratively modified and its sensibility evaluated by field-testing it on children with LD, and their parents. Cognitive interviews were conducted with a subgroup of these children, and an e-survey administered to a multidisciplinary group of health care professionals with expertise in paediatric LD. Findings were integrated to create the final version of the GOAL-LD. Twenty-five children (9-18 years), 20 parents, and 31 healthcare professionals evaluated the content and sensibility of the GOAL, with an emphasis on the relevance and importance of the items to patients' health related quality of life (HRQL). This resulted in the retention of 26 of the original 50 items, elimination of 12, modification of 12, and addition of seven new items. The new 45-item GOAL-LD questionnaire was shown to be sensible, and its content deemed important. The GOAL-LD questionnaire has a high level of face and content validity, and sensibility. It comprehensively captures the HRQL goals and outcomes that matter to children with LD and their parents. Following further psychometric evaluation, the GOAL-LD may serve as a much needed patient and parent reported outcome measure for this population.


TCF3 mutually exclusive alternative splicing is controlled by long-range cooperative actions between hnRNPH1 and PTBP1.
TCF3, also known as E2A, is a well-studied transcription factor that plays an important role in stem cell maintenance and hematopoietic development. The TCF3 gene encodes two related proteins, E12 and E47, which arise from mutually exclusive alternative splicing (MEAS). Since these two proteins have different DNA binding and dimerization domains, this AS event must be strictly regulated to ensure proper isoform ratios. Previously, we found that heterogeneous nuclear ribonucleoprotein (hnRNP) H1/F regulates TCF3 AS by binding to exonic splicing silencers (ESSs) in exon 18b. Here, we identify conserved intronic splicing silencers (ISSs) located between, and far from, the two mutually exclusive exons, and show that they are essential for MEAS. Further, we demonstrate that the hnRNP PTBP1 binds the ISS and is a regulator of TCF3 AS. We also demonstrate that hnRNP H1 and PTBP1 regulate TCF3 AS reciprocally, and that position-dependent interactions between these factors are essential for proper TCF3 MEAS. Our study provides a new model in which MEAS is regulated by cooperative actions of distinct hnRNPs bound to ISSs and ESSs.


Nitrite Activates 5'AMP-Activated Protein Kinase-Endothelial Nitric Oxide Synthase Pathway in Human Glomerular Endothelial Cells.
Recent studies have shown that orally supplied nitrates, which substantially exist in our daily diets, are reduced into nitrites and become significant sources of nitric oxide (NO) especially in hypoxic tissues. However, physiological significance of nitrites in normal tissues has not been elucidated though our serum concentrations of nitrites reach as high as micromolar levels. We investigated effects of nitrite on endothelial NO synthase (eNOS) using human glomerular endothelial cells to reveal potential glomerular-protective actions of nitrites with its underlying molecular mechanism. Here we demonstrate that nitrite stimulation evokes eNOS activation which is dependent on 5'AMP-activated protein kinase (AMPK) activation in accordance with ATP reduction. Thus, nitrites should facilitate AMPK-eNOS pathway in an energy level-dependent manner in endothelial cells. The activation of AMPK-eNOS signals is suggested to be involved in vascular and renal protective effects of nitrites and nitrates. Nitrites may harbor beneficial effects on metabolic regulations as AMPK activators.


Percutaneous cholecystostomy (PC) in the management of acute cholecystitis in high risk patients.
To determine the role of ultrasound-guided percutaneous cholecystostomy (PC) regarding complications and outcome in the management of acute cholecystitis in patients high risk for surgery and anaesthesia and not responding to conservative management. Observational case series. The study was carried out at The Aga Khan University Hospital, Karachi, from January 2003 to December 2007. The study included patients admitted with acute cholecystitis considered unfit for immediate surgery but not responding to conservative management. Percutaneous cholecystostomy was conducted under ultrasound guidance. The studied variables included patients' demographics, co-morbid, ultrasound findings of biliary tree, indication for percutaneous cholecystostomy, its route, complication during or after procedure, patient's clinical outcome (upto 48 hours) and 30 days follow-up. Those with incomplete medical record and follow-up were excluded. Data were analyzed and results compiled using SPSS 16.0 version. Mean and standard deviation for quantitative variable like age was derived. Proportions were computed for complications and patient's clinical outcome. Forty one patients with complete medical record were studied including 15 (37%) males and 26 (63%) females. Mean age was 65 +/-13.5 years. Indications for PC included calculus cholecystitis in 25, acalculous cholecystitis in 10, empyema in 04 and gallbladder perforation in 02 patients. No complication was seen during or after procedure in 31 (75%) patients. Complications occurred in 10 (25%) patients including vagal reaction, pain during procedure, tube blockage, catheter dislodgement and bile leakage. Favourable clinical response (improvement in clinical symptoms) was noted in 34 (83%) patients. Seven (17%) patients did not show any improvement in clinical condition after the procedure. On 30 days follow-up, 9 patients had undergone cholecystectomy, 5 (12%) patients expired due to underlying clinical conditions and the rest were settled without requiring an immediate cholecystectomy. There was no direct procedure-related mortality. Imaging guided PC is a safe and effective procedure for immediate management of non-resolving acute cholecystitis in patients high risk for surgery and anaesthesia and not responding to conservative management.


The Elevated High-Sensitivity Cardiac Troponin T Pilot: Diagnoses and Outcomes.
To identify the diagnoses and outcomes associated with elevated high sensitivity cardiac troponin T (hs-cTnT) compared with the 4th-generation troponin T and to validate the Mayo Clinic hs-cTnT myocardial infarction algorithm cutoff values. Consecutive blood samples of patients presenting to the emergency department between July 2017 and August 2017, who had 4th-generation troponin T, were also analyzed using the hs-cTnT assay. Troponin T values, discharge diagnoses, comorbidities, and outcomes were assessed. In addition, analyses of sex-specific and hs-cTnT cutoff values were assessed. Of 830 patients, 32% had an elevated 4th-generation troponin T, whereas 64% had elevated hs-cTnT. With serial sampling, 4th-generation troponin missed a chronic myocardial injury pattern and acute myocardial injury pattern in 64% and 16% of patients identified with hs-cTnT, respectively. Many of these "missed" patients had discharge diagnoses associated with cardiovascular disease, infection, or were postoperative. Five of the 6 patients with unstable angina ruled in for myocardial infarction. There were many increases in hs-cTnT that were missed by the 4th-generation cTnT assay. Most new increases are not related to acute cardiac causes. They were more consistent with chronic myocardial injury. High-sensitivity cTnT did reclassify most patients with unstable angina as having non-ST-elevation myocardial infarction. Older age, more comorbidities, and lower hemoglobin were associated with elevated hs-cTnT. Our data also support the use of our sex-specific cutoff values.


Evaluation of animal models for the study of exercise-induced muscle enlargement.
Skeletal muscle is known to enlarge in response to high-resistance training programs in humans. Study of the cellular mechanisms of muscle enlargement and the adaptations of muscle to strength-training programs has been difficult because of the need to analyze entire muscles. This precludes the use of human subjects in many experiments of this nature. Several animal models have been developed for the study of muscle enlargement; these models basically fall into three categories: 1) stretch hypertrophy, 2) compensatory hypertrophy, and 3) exercise-induced hypertrophy. This review attempts to analyze these models as models of muscle enlargement produced by strength training in humans. Three areas must be considered when evaluating animal models of human muscle enlargement produced by strength training: 1) response topography, 2) magnitude of enlargement, and 3) muscle fiber adaptations produced as a result of the enlargement. Based on these considerations, it is concluded that none of the animal models currently in use truly represents the human strength-training situation under all conditions. All three models, however, provide valuable information about the plasticity of skeletal muscle in response to a broad spectrum of muscle enlargement.


The chemokines secretion and the oxidative stress are targets of low-level laser therapy in allergic lung inflammation.
Recent studies show that low-level laser therapy (LLLT) has an important anti-inflammatory action in acute lung inflammation. The present work explored if laser therapy is able to antagonize eosinophils and allergic inflammation induced by oxidative stress in Balb/c mice. Forty-eight hours after challenge, the leukocyte counting, ROS and nitrite/nitrate level, RANTES, CCL3, CCL8 as well as eotaxins were measured in the bronchoalveolar lavage fluid (BALF) of laser-treated mice or not. Into the lung, some chemokines receptors, the iNOS activity and mRNA expression, and the activities of superoxide dismutase (SOD), catalase, gluthatione, NADPH oxidase activities and thiobarbituric acid reactive species (T-Bars) were measured. Laser-treated allergic mice presented reduction of both the ICAM-1 and eosinophil in the lungs. RANTES, CCL8, CCL3 and eotaxins were reduced in BALF of laser-treated allergic mice. In allergic mice lung LLLT decreased the CCR1 and CCR3 and restored the oxidative stress balance as well. Laser decreased the lipidic peroxidation in allergic mice lung as much as increased SOD, GPx and GR. It shows that LLLT on allergic lung inflammation involves leukocyte-attractant chemokines and endogenous antioxidant. Based on results, LLLT may ultimately become a non- invasive option in allergic lung disease treatment. The top figure illustrates the laser decreasing the eosinophils migration into BALF and the bottom figure shows the laser upregulating the expression of heme-oxygenase (anti-oxidant enzyme) in lung tissue anti-oxidant.


A study on hydrogen storage performance of Ti decorated vacancies graphene structure on the first principle.
The structural properties, formation energy, adsorption energy, and electronic properties of vacancy graphene are studied by first-principles analysis. We found that the formation energy and adsorption energy of double vacancy graphene (DVG-4) are the largest. A single defect in DVG-4 can adsorb at least nine hydrogen molecules, and compared with Ti modified single vacancy graphene (SVG-Ti), the adsorption capacity is increased by 80%. When DVG-4 adsorbs the second, third, and fourth hydrogen molecules, the adsorption energy is greater than 0.7 eV, which is not conducive to the release. Density of state (DOS) and electron density difference (EDIFF) results reveal that charge transfer occurs among hydrogen molecules, Ti atoms, and DVG-4, decreasing the hydrogen adsorption capacity of DVG-4 by 33%. DVG - 4 has the potential to become an excellent hydrogen storage material.


Commitment to nutritional endoderm in Eleutherodactylus coqui involves altered nodal signaling and global transcriptional repression.
The vegetal cells of a Xenopus laevis embryo commit to mesendoderm via the Nodal-signaling pathway. In the direct developing frog Eleutherodactylus coqui, mesendoderm is specified at the marginal zone of the early gastrula, and vegetal core cells transform into nutritional endoderm. Nutritional endoderm, a novel tissue, consists of transient, yolky cells that provide nutrition but remain undifferentiated. We report a dual regulation for the generation of nutritional endoderm. First, differential expressions of the Nodal-signal transducers Smad2 and Smad4 were observed during early gastrulation between the marginal zone and the vegetal core cells. Although EcSmad2 RNA as well as total and activated Smad2 protein were detected in the vegetal core, Smad4 protein was expressed less in vegetal core during early gastrulation. Only 12% and 50% of vegetal core cells were positive for nuclear Smad2 and Smad4 signals respectively compared to 100% of marginal zone cells. These results suggest a signaling disruption in the vegetal core. Second, vegetal core cells were transcriptionally repressed. At the blastula stage, both marginal zone and vegetal core cells were transcriptionally silent, but during early gastrulation, only marginal zone cells became transcriptionally active. This indicates the occurrence of a mid-blastula transition in the marginal zone by early gastrulation, but global transcriptional repression persisted in the vegetal core and its derivative, nutritional endoderm, throughout development. We have described a novel mechanism, which prevents differentiation of the vegetal core through differential Nodal-signaling and global transcriptional repression.


A qualitative assessment of factors affecting nursing home caregiving staff experiences during the COVID-19 pandemic.
A large portion of COVID-19 cases and deaths in the United States have occurred in nursing homes; however, current literature including the frontline perspective of staff working in nursing homes is limited. The objective of this qualitative assessment was to better understand what individual and facility level factors may have contributed to the impact of COVID-19 on Certified Nursing Assistants (CNAs) and Environmental Services (EVS) staff working in nursing homes. Based on a simple random sample from the National Healthcare Safety Network (NHSN), 7,520 facilities were emailed invitations requesting one CNA and/or one EVS staff member for participation in a voluntary focus group over Zoom. Facility characteristics were obtained via NHSN and publicly available sources; participant demographics were collected via SurveyMonkey during registration and polling during focus groups. Qualitative information was coded using NVIVO and Excel. Throughout April 2021, 23 focus groups including 110 participants from 84 facilities were conducted homogenous by participant role. Staffing problems were a recurring theme reported. Participants often cited the toll the pandemic took on their emotional well-being, describing increased stress, responsibilities, and time needed to complete their jobs. The lack of consistent and systematic guidance resulting in frequently changing infection prevention protocols was also reported across focus groups. Addressing concerns of low wages and lack of financial incentives may have the potential to attract and retain employees to help alleviate nursing home staff shortages. Additionally, access to mental health resources could help nursing home staff cope with the emotional burden of the COVID-19 pandemic. These frontline staff members provided invaluable insight and should be included in improvement efforts to support nursing homes recovering from the impact of COVID-19 as well as future pandemic planning.


Comparing a Rule Based vs. Statistical System for Automatic Categorization of MEDLINE Documents According to Biomedical Specialty.
Automatic document categorization is an important research problem in Information Science and Natural Language Processing. Many applications, including Word Sense Disambiguation and Information Retrieval in large collections, can benefit from such categorization. This paper focuses on automatic categorization of documents from the biomedical literature into broad discipline-based categories. Two different systems are described and contrasted: CISMeF, which uses rules based on human indexing of the documents by the Medical Subject Headings(®) (MeSH(®)) controlled vocabulary in order to assign metaterms (MTs), and Journal Descriptor Indexing (JDI) based on human categorization of about 4,000 journals and statistical associations between journal descriptors (JDs) and textwords in the documents. We evaluate and compare the performance of these systems against a gold standard of humanly assigned categories for one hundred MEDLINE documents, using six measures selected from trec_eval. The results show that for five of the measures, performance is comparable, and for one measure, JDI is superior. We conclude that these results favor JDI, given the significantly greater intellectual overhead involved in human indexing and maintaining a rule base for mapping MeSH terms to MTs. We also note a JDI method that associates JDs with MeSH indexing rather than textwords, and it may be worthwhile to investigate whether this JDI method (statistical) and CISMeF (rule based) might be combined and then evaluated showing they are complementary to one another.


Two-Dimensional Penta-BN2 with High Specific Capacity for Li-Ion Batteries.
Searching for high-performance electrode materials is one of the most effective ways to improve the energy density of current lithium-ion batteries. Using first-principles calculations, we reveal that pentagonal BN2 (Penta-BN2) can serve as a compelling anode material for lithium-ion batteries. Penta-BN2 harbors intrinsic metallic nature before and after lithiation, showing excellent electrical conductivity. Significantly, the fully lithiated phase of Penta-BN2 is Li3BN2, corresponding to an ultra-high theoretical capacity of 2071 mAh g-1, superior to the capacity of most of the previously reported two-dimensional candidates. Meanwhile, the open-circuit voltage and diffusion barrier height of Penta-BN2 are found to be fascinatingly low. Moreover, in light of its small Young's modulus and robust lattice toward lithiation, Penta-BN2 can accommodate volume changes during the charging-discharging processes, remarkably beneficial for fabricating flexible electrodes.


Age, cumulative trauma and stressful life events, and post-traumatic stress symptoms among older adults in prison: do subjective impressions matter?
The aging prison population in the United States presents a significant public health challenge with high rates of trauma and mental health issues that the correctional system alone is ill-prepared to address. The purpose of this study was to examine the relationship of age, objective, and subjective measures of trauma and stressful life events and post-traumatic stress symptoms among older adults in prison. Data were gathered from 334 prisoners (aged 55+) housed in the New Jersey Department of Corrections, as of September 2010. An anonymous self-report, self-administered survey was mailed to the total population of 1,000 prisoners aged 55 years and older. Objective and subjective trauma was measured using the Life Stressors Checklist-Revised (LSC-R), and post-traumatic stress symptoms were measured using the Civilian Version of the Post-traumatic Stress Scale. Results of a path analysis revealed that past year subjective impressions of traumatic and stressful life events had a positive and significant relationship to current post-traumatic stress symptoms. Age was found to have a significant and inverse relationship to subjective traumatic and stressful life events. That is, younger participants reported higher levels of cumulative traumatic and stressful life events and past year subjective ratings of being bothered by these past events. These findings have significance for interdisciplinary/interprofessional practice and appropriate institutional and community care, including reentry planning of older adults in the criminal justice system.


Placental expression of pituitary hormones is an ancestral feature of therian mammals.
The placenta is essential for supplying nutrients and gases to the developing mammalian young before birth. While all mammals have a functional placenta, only in therian mammals (marsupials and eutherians) does the placenta closely appose or invade the uterine endometrium. The eutherian placenta secretes hormones that are structurally and functionally similar to pituitary growth hormone (GH), prolactin (PRL) and luteinizing hormone (LH). Marsupial and eutherian mammals diverged from a common ancestor approximately 125 to 148 million years ago and developed distinct reproductive strategies. As in eutherians, marsupials rely on a short-lived but functional placenta for embryogenesis. We characterized pituitary GH, GH-R, IGF-2, PRL and LHβ in a macropodid marsupial, the tammar wallaby, Macropus eugenii. These genes were expressed in the tammar placenta during the last third of gestation when most fetal growth occurs and active organogenesis is initiated. The mRNA of key growth genes GH, GH-R, IGF-2 and PRL were expressed during late pregnancy. We found significant up-regulation of GH, GH-R and IGF-2 after the start of the rapid growth phase of organogenesis which suggests that the placental growth hormones regulate the rapid phase of fetal growth. This is the first demonstration of the existence of pituitary hormones in the marsupial placenta. Placental expression of these pituitary hormones has clearly been conserved in marsupials as in eutherian mammals, suggesting an ancestral origin of the evolution of placental expression and a critical function of these hormones in growth and development of all therian mammals.


A huge staghorn renal stone: Is there still a need for open surgery to protect against further damage to the kidney? A case report.
Staghorn calculi (SC) occupy the renal pelvis and calyces and are common in females linked to repeated urinary tract infections (UTIs). Judicious surgery planning reduces the chance of further damage to the kidney due to renal SCs. Open stone surgery (OSS) is one of the various operative techniques to remove such huge stones with one operative intervention and protect the kidney from ongoing functional damage. A 47-year-old male patient presented with right-sided renal colic pain, and on further investigations, he was diagnosed with a large renal stone responsible for substantial renal function impairment on the same side. The SC measured 8 × 4 cm with another stone in the lower calyx. thinner parenchyma, and only 16% relative function. Therefore, open surgery was selected over less invasive approaches because multiple lithotripsy (ESWL) sittings may have been required in less invasive options. SCs, which can be complete or partial, often result in renal impairment. Hence, it is crucial to implement a proactive therapeutic approach that includes a thorough evaluation of the stone's size and position, the patient's choice, and institutional capacity. Complete elimination of SCs is preferred to maintain maximal renal function. Based on clinical, technical, and socioeconomic considerations, open pyelolithotomy or OSS was chosen over percutaneous nephrolithotomy for SC removal in the discussed case. The ability to remove large stones in a single intervention with open pyelolithotomy has been very effective due to its distinctive clinical presentation and pathological abnormalities.


Evolutionary influences in arboviral disease.
Arthropod-borne viruses (arboviruses) generally require horizontal transmission by arthropod vectors among vertebrate hosts for their natural maintenance. This requirement for alternate replication in disparate hosts places unusual evolutionary constraints on these viruses, which have probably limited the evolution of arboviruses to only a few families of RNA viruses (Togaviridae, Flaviviridae, Bunyaviridae, Rhabdoviridae, Reoviridae, and Orthomyxoviridae) and a single DNA virus. Phylogenetic studies have suggested the dominance of purifying selection in the evolution of arboviruses, consistent with constraints imposed by differing replication environments and requirements in arthropod and vertebrate hosts. Molecular genetic studies of alphaviruses and flaviviruses have also identified several mutations that effect differentially the replication in vertebrate and mosquito cells, consistent with the view that arboviruses must adopt compromise fitness characteristics for each host. More recently, evidence of positive selection has also been obtained from these studies. However, experimental model systems employing arthropod and vertebrate cell cultures have yielded conflicting conclusions on the effect of alternating host infections, with host specialization inconsistently resulting in fitness gains or losses in the bypassed host cells. Further studies using in vivo systems to study experimental arbovirus evolution are critical to understanding and predicting disease emergence, which often results from virus adaptation to new vectors or amplification hosts. Reverse genetic technologies that are now available for most arbovirus groups should be exploited to test assumptions and hypotheses derived from retrospective phylogenetic approaches.


Fasciola hepatica miracidia: lectin binding and stimulation of in vitro miracidium-to-sporocyst transformation.
The lectin binding properties of Fasciola hepatica miracidia were studied by a panel of fluorescein- and gold-conjugated lectins (ConA, LCA, WGA, LEA, SBA, HPA and UEA-I). The presence of mannose and/or glucose residues was demonstrated with ConA and LCA as weak diffuse fluorescence of the miracidial surface, which was more intense at the anterior part of the larva. The N-acetylglucosamine-binding lectins WGA and LEA reacted intensely with the whole miracidial surface. No labelling with N-acetylgalactosamine and/or galactose-specific (SBA and HPA) and fucose-specific UEA-I lectins was observed. The possibility that the specific recognition of the miracidial surface carbohydrates by lectins may initiate the process of transformation of the miracidia into sporocysts was examined in vitro in physiological saline for Galba truncatula. Incubation in the presence of ConA and WGA resulted in facilitation of the transformation process. Facilitation was absent in the presence of inhibitor sugars. Incubation in the presence of SBA or UEA-I had no effect. The results suggested a possible impact of carbohydrate-lectin interactions in transformation of miracidia of F. hepatica to sporocysts in vivo.


Does the addition of tramadol and ketamine to ropivacaine prolong the axillary brachial plexus block?
A prospective, randomized, controlled, double-blind clinical trial to assess the effect of tramadol and ketamine, 50 mg, added to ropivacaine in brachial plexus anesthesia. Thirty-six ASA physical statuses I and II patients, between 18 and 60 years of age, scheduled for forearm and hand surgery under axillary brachial plexus block, were allocated to 3 groups. Group R received 0.375% ropivacaine in 40 mL, group RT received 0.375% ropivacaine in 40 mL with 50 mg tramadol, and group RK received 0.375% ropivacaine in 40 mL with 50 mg ketamine for axillary brachial plexus block. The onset times and the duration of sensory and motor blocks, duration of analgesia, hemodynamic parameters, and adverse events (nausea, vomiting, and feeling uncomfortable) were recorded. The onset time of sensorial block was the fastest in ropivacaine + tramadol group. Duration of sensorial and motor block was the shortest in the ropivacaine + tramadol group. Duration of analgesia was significantly longer in ropivacaine + tramadol group. We conclude that when added to brachial plexus analgesia at a dose of 50 mg, tramadol extends the onset and duration time of the block and improves the quality of postoperative analgesia without any side effects.


Activation of the erythropoietin receptor in human skeletal muscle.
Erythropoietin receptor (EPOR) expression in non-hematological tissues has been shown to be activated by locally produced and/or systemically delivered EPO. Improved oxygen homeostasis, a well-established consequence of EPOR activation, is very important for human skeletal muscle performance. In the present study we investigate whether human skeletal muscle fibers and satellite cells express EPOR and if it is activated by exercise. Ten healthy males performed 65 min of cycle exercise. Biopsies were obtained from the vastus lateralis muscle and femoral arterio-venous differences in EPO concentrations were estimated. The EPOR protein was localized in areas corresponding to the sarcolemma and capillaries. Laser dissection identified EPOR mRNA expression in muscle fibers. Also, EPOR mRNA and protein were both detected in human skeletal muscle satellite cells. In the initial part of the exercise bout there was a release of EPO from the exercising leg to the circulation, possibly corresponding to an increased bioavailability of EPO. After exercise, EPOR mRNA and EPOR-associated JAK2 phosphorylation were increased. Interaction with JAK2 is required for EPOR signaling and the increase found in phosphorylation is therefore closely linked to the activation of EPOR. The receptor activation by acute exercise suggests that signaling through EPOR is involved in exercise-induced skeletal muscle adaptation, thus extending the biological role of EPO into the skeletal muscle.


A Novel Modelling Methodology Which Predicts the Structural Behaviour of Vertebral Bodies under Axial Impact Loading: A Finite Element and DIC Study.
Cervical spine injuries (CSIs) arising from collisions are uncommon in contact sports, such as rugby union, but their consequences can be devastating. Several FE modelling approaches are available in the literature, but a fully calibrated and validated FE modelling framework for cervical spines under compressive dynamic-impact loading is still lacking and material properties are not adequately calibrated for such events. This study aimed to develop and validate a methodology for specimen-specific FE modelling of vertebral bodies under impact loading. Thirty-five (n = 35) individual vertebral bodies (VBs) were dissected from porcine spine segments, potted in bone cement and μCT scanned. A speckle pattern was applied to the anterior faces of the bones to allow digital image correlation (DIC), which monitored the surface displacements. Twenty-seven (n = 27) VBs were quasi-statically compressively tested to a load up to 10 kN from the cranial side. Specimen-specific FE models were developed for fourteen (n = 14) of the samples in this group. The material properties were optimised based on the experimental load-displacement data using a specimen-specific factor (kGSstatic) to calibrate a density to Young's modulus relationship. The average calibration factor arising from this group was calculated (K¯GSstatic) and applied to a control group of thirteen (n = 13) samples. The resulting VB stiffnesses was compared to experimental findings. The final eight (n = 8) VBs were subjected to an impact load applied via a falling mass of 7.4kg at a velocity of 3.1ms-1. Surface displacements and strains were acquired from the anterior VB surface via DIC, and the impact load was monitored with two load cells. Specimen-specific FE models were created for this dynamic group and material properties were assigned again based on the density-Young's modulus relationship previously validated for static experiments, supplemented with an additional factor (KGSdynamic). The optimised conversion factor for quasi-static loading, K¯GSstatic, had an average of 0.033. Using this factor, the validation models presented an average numerical stiffness value 3.72% greater than the experimental one. From the dynamic loading experiments, the value for KGSdynamic was found to be 0.14, 4.2 times greater than K¯GSstatic. The average numerical stiffness was 2.3% greater than in the experiments. Almost all models presented similar stiffness variations and regions of maximum displacement to those observed via DIC. The developed FE modelling methodology allowed the creation of models which predicted both static and dynamic behaviour of VBs. Deformation patterns on the VB surfaces were acquired from the FE models and compared to DIC data, achieving high agreement. This methodology is now validated to be fully applied to create whole cervical spine models to simulate axial impact scenarios replicating rugby collision events.