Functional connectivity of the human insular cortex during noxious and innocuous thermal stimulation.
The insula plays a key role in brain processing of noxious and innocuous thermal stimuli. The anterior and the posterior portions of the insular cortex are involved in different ways in nociceptive and thermoceptive processing. Therefore, their stimulus-specific functional connectivity may also differ. Here we used functional magnetic resonance imaging (fMRI) to investigate the activity and functional connectivity of insular cortex subregions during noxious and innocuous thermal stimulation. In 11 healthy subjects, psychophysically controlled noxious and innocuous warm and cold stimuli were applied to the left forearm. To differentiate between the subregions of the insular cortex involved in pain processing and those involved in temperature processing, a 2×2 factorial fMRI analysis was performed. Pain processing insular areas (main effect of pain) were detected in bilateral aINS and contralateral pINS. Temperature processing insular areas (main effect of temperature) were also found in bilateral aINS and contralateral pINS. The individual signal time courses from the pain- and temperature processing insular activation clusters were used for calculation and comparison of stimulus-specific functional connectivity of aINS and pINS by means of a correlation analysis. As expected, both aINS and pINS were functionally connected to a large brain network - which predominantly includes areas involved in nociception and thermoception: primary (S1) and secondary (S2) somatosensory cortices, cingulate gyrus, prefrontal cortex (PFC) and parietal association cortices (PA). When statistically compared, during both noxious and innocuous stimulation, aINS was more strongly connected to PFC and to ACC than was pINS; pINS meanwhile was more strongly connected to S1 and to the primary motor cortex (M1). Interestingly, S2 was more strongly connected to aINS than to pINS during painful stimulation but not during innocuous thermal stimulation. We conclude that aINS is more strongly functionally connected to areas known for affective and cognitive processing, whereas pINS is more strongly connected with areas known for sensory-discriminative processing of noxious and somatosensory stimuli.


General anesthesia alters the diversity and composition of the lung microbiota in rat.
The lung microbiome plays a crucial role in human health and disease. Extensive studies have demonstrated that the disturbance of the lung microbiome influences immune response, cognition, and behavior. The goal of this study was to investigate the effect of general anesthetics on lung microbiome. Eight-week-old male SD rats received a continuous intravenous infusion of propofol or inhalation of isoflurane for 4 h. 16S rRNA gene amplification from BALF samples was used to investigate the changes in the lung microbiome after interventions. We further performed neurobehavioral assessments to find the differential strains' association with behavior disorder after isoflurane anesthesia. The absolute and relative quantitation of 16S rRNA sequencing data showed that isoflurane altered the diversity and abundance of the lung microbiome in rats more than propofol. Elusimicrobia increased significantly in the isoflurane group. Both EPM and OFT results showed that rats exhibited depression-like behaviors after inhalation of isoflurane. In addition, significant differences were found in the COG/KO/MetaCyc/KEGG pathway enrichment analyses among the groups. Continuous inhalation of isoflurane changed the diversity and composition of the lung microbiota in rats, resulting in post-anesthesia depression.


Adding liposomal doxorubicin enhances the abscopal effect induced by radiation/αPD1 therapy depending on tumor cell mitochondrial DNA and cGAS/STING.
Localized radiotherapy (RT) can cause a T cell-mediated abscopal effect on non-irradiated tumor lesions, especially in combination with immune checkpoint blockade. However, this effect is still clinically rare and improvements are highly desirable. We investigated whether triple combination with a low dose of clinically approved liposomal doxorubicin (Doxil) could augment abscopal responses compared with RT/αPD-1 and Doxil/αPD-1. We also investigated whether the enhanced abscopal responses depended on the mitochondrial DNA (mtDNA)/cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING)/IFN-I pathway. We used Doxil in combination with RT and αPD-1 in two tumor models (B16-CD133 melanoma and MC38 colon carcinoma) with mice bearing two tumors, only one of which was irradiated. Mechanistic studies on the role of the mtDNA/cGAS/STING/IFN-I axis were performed using inhibitors and knockout cells in vitro as well as in mice. Addition of a single low dose of Doxil to RT and αPD-1 strongly enhanced the RT/αPD-1-induced abscopal effect in both models. Complete cures of non-irradiated tumors were mainly observed in triple-treated mice. Triple therapy induced more cross-presenting dendritic cells (DCs) and more tumor-specific CD8+ T cells than RT/αPD-1 and Doxil/αPD-1, particularly in non-irradiated tumors. Coincubation of Doxil-treated and/or RT-treated tumor cells with DCs enhanced DC antigen cross-presentation which is crucial for inducing CD8+ T cells. CD8+ T cell depletion or implantation of cGAS-deficient or STING-deficient tumor cells abolished the abscopal effect. Doxorubicin-induced/Doxil-induced IFNβ1 markedly depended on the cGAS/STING pathway. Doxorubicin-treated/Doxil-treated tumor cells depleted of mtDNA secreted less IFNβ1, of the related T cell-recruiting chemokine CXCL10, and ATP; coincubation with mtDNA-depleted tumor cells strongly reduced IFNβ1 secretion by DCs. Implantation of mtDNA-depleted tumor cells, particularly at the non-irradiated/abscopal site, substantially diminished the Doxil-enhanced abscopal effect and tumor infiltration by tumor-specific CD8+ T cells. These data show that single low-dose Doxil can substantially enhance the RT/αPD-1-induced abscopal effect, with a strong increase in cross-presenting DCs and CD8+ tumor-specific T cells particularly in abscopal tumors compared with RT/αPD-1 and Doxil/αPD-1. Moreover, they indicate that the mtDNA/cGAS/STING/IFN-I axis is important for the immunogenic/immunomodulatory doxorubicin effects. Our findings may be helpful for the planning of clinical radiochemoimmunotherapy trials in (oligo)metastatic patients.


Erratum: Effect of Breakfast Consumption and Meal Time Regularity on Nutrient Intake and Cardiometabolic Health in Korean Adults.
[This corrects the article on p. 240 in vol. 10, PMID: 34095015.].


Adverse effects of pesticides on biochemical and haematological parameters in Tunisian agricultural workers.
Biomonitoring of effects in agricultural workers is necessary to assess the individual risk of handling pesticides. In this study, biochemical and haematological parameters were measured to evaluate the effects of exposure to these compounds in agricultural workers. The study was carried out in 110 workers and 97 control subjects. Several haematological and biochemical parameters were analysed. Assessment of haematological parameters revealed that the mean cell volume and haematocrit levels were significantly lower in workers than in controls (P=0.002 and 0.013, respectively), while mean corpuscular haemoglobin concentrations were higher in workers (P<0.001). There was also a significant inhibition of butyrylcholinesterase activity in workers compared with that in controls (P<0.001). Assessment of biochemical parameters further showed significantly higher activities of transferases, lactate dehydrogenase (P<0.001), alkaline phosphatase (ALP) (P=0.006) and creatine kinase (CK) (P<0.015), as well as higher levels of proteins (P<0.001), creatinine (P=0.001) and urea (P=0.001) in workers compared with controls, along with significantly higher uric acid levels (P=0.012). Furthermore, the number of years exposed to pesticides predicted higher activities of alanine aminotransferase, CK, ALP, as well as uric acid levels. Overall, chronic exposure to pesticides appeared to affect several biochemical parameters. These biomarkers seem to be indicative of adverse effects of pesticides in agricultural workers, confirming their use for routine monitoring of effects.


Grey matter volume and its association with cognitive impairment and peripheral cytokines in excited individuals with schizophrenia.
Although excited behavior in patients with schizophrenia has been linked to brain structural abnormalities, whether cortical abnormalities in this subgroup are related to cognitive impairment or peripheral immune responses is unknown. We included 28 patients with excitability (EC), 28 patients without excitability (NEC), and 48 healthy controls (HCs) to evaluate the associations. Compared with the HC group, the EC and NEC groups showed significant cognitive impairment and increased serum cytokine levels. Analysis of variance in whole-brain grey matter volume (GMV) showed that the volumes of several brain areas, including the superior frontal gyrus (SFG), superior temporal gyrus, and cingulate gyrus, were decreased in patients with schizophrenia. Notably, the left SFG volume was significantly lower in the EC group than in the NEC group. Spearman correlation analysis showed that elevated cytokines were negatively correlated with the GMV of the bilateral SFG, bilateral inferior parietal gyrus, left anterior cingulate gyrus, right fusiform gyrus and parahippocampal gyrus, which were mainly positive correlated with cognitive tests. Moreover, interleukin 4 may contribute to poor scores on Brief Assessment of Cognition and Neuropsychological Assessment Battery by reducing the left SFG volume (17%, Pmediation = 0.044; and 24%, Pmediation = 0.040, respectively). In conclusion, our results confirm GMV changes in excited patients with schizophrenia, and characterize the GMV as an important interface between inflammatory cytokines and cognitive impairment. Therefore, targeted anti-inflammatory adjuvant antipsychotic therapy may improve cognitive function and volumetric brain abnormalities in these patients.


A comparison of maintenance of canal curvature using balanced-force instrumentation with three different file types.
Forty-five extracted human molar teeth with curved facial or mesial canals were instrumented to a #45 file with three different file types using balanced-force (BF) instrumentation. Pre- and postoperative X-rays were taken, projected, and file location traced to determine the canal curvature according to the method of Schneider. Results showed statistically less reduction in canal curvature with nickel-titanium (NT) files compared with either Flex-R (FR) (p < 0.05) or K-Flex (KF) files (p < 0.0021). There was no significant difference comparing FR with KF files (p > 0.05). Under the conditions of this study, NT files used with the BF technique were significantly less likely to change canal curvature than either FR or KF files when instrumenting curved root canals.


Qualitative comparison of curricula in oral and maxillofacial surgery training. Part 2: oral surgery.
A comparison between the curricula for specialist training in oral surgery (OS), and oral and maxillofacial surgery (OMFS) illustrates the overlap between the two specialties. We identified and compared relevant curricula for OMFS with competencies in the OS specialist-training curriculum using a qualitative assessment to establish the degree of overlap. All competencies within the OS curriculum are covered by OMFS curricula, and 21 of 96 OMFS clinical competencies are covered by OS core competencies. The majority of OMFS competencies are unique and are not directly comparable with those in the OS curriculum. Knowledge of this overlap may help to avoid unnecessary duplication in the training of those who wish to transfer specialty.


Spectroscopic properties of siroheme extracted from sulfite reductases.
Siroheme has been extracted from sulfite reductases and its properties in aqueous solution have been investigated by optical absorption, electron paramagnetic resonance (EPR), and magnetic circular dichroism (MDC) spectroscopy. The absorption spectrum of siroheme exhibits a marked pH dependence, and two pK values, 4.2 and 9.0, were determined by pH titration in the range 2-12. The first pK (4.2) is thought to correspond to the ionization of the carboxylic acid side-chains on the tetrapyrrole rings, and the second pK (9.0) is attributed to displacement of the axial ligand chloride by hydroxide. The binding of the strong field ligands, CO, NO, and cyanide, were investigated by UV-visible absorption and, in the case of the cyanide complex, by low-temperature EPR and MCD spectroscopies. CO and NO were able to reduce and bind to siroheme without additional reducing agent. The EPR spectrum of the isolated siroheme (chloride-ferrisiroheme) exhibits an axial signal with g perpendicular = 6.0 and g parallel = 2.0, typical of high-spin ferric hemes (S = 5/2), whereas the cyanide-complexed siroheme exhibits an approximately axial signal with g perpendicular = 2.38 and g parallel = 1.76 that is indicative of a low-spin ferric heme (S = 1/2). The low-temperature MCD spectra and magnetization data for the as-isolated and cyanide-complexed ferrisiroheme are entirely consistent with the interpretation of the EPR spectra. The results for ferrosiroheme indicate that the siroheme remains high spin (S = 2) and low spin (S = 0) on reduction of the as-isolated and cyanide-complexed siroheme, respectively. The isolated siroheme expressed sulfite reductase activity but the assessable catalytic cycle was much less than that of the native enzyme, showing the importance of the protein environment.


Influence of intermittent compressive force on proteoglycan content in calcifying growth plate cartilage in vitro.
We investigated the effect of mechanical stimulation by an intermittent compressive force (ICF) on proteoglycan (PG) synthesis and PG structure in calcified and noncalcified cartilage of fetal mouse long bone rudiments. Uncalcified cartilaginous long bone rudiments were cultured for 5 days in the presence of [35S]sulfate and [3H]glucosamine under control conditions (atmospheric pressure) or under the influence of ICF. ICF was generated by intermittently compressing the gas phase above the culture medium (130 mbar, 0.3 Hz). During culture, the center of the rudiments started to calcify. ICF stimulated calcification such that, after 5 days, the diaphysis of calcified cartilage was about two times as long as in the control cultures. At the end of the experiment, the rudiments were divided in a central calcified diaphysis and two noncalcified epiphyses. Diaphysis and epiphyses were pooled separately. PGs were extracted with 4 M guanidinium chloride and isolated by cesium chloride density gradient centrifugation. PGs (predigested with proteinase K or chondroitinase ABC) were characterized for hydrodynamic size of aggregates, monomers, and chondroitin sulfate chains by gel permeation chromatography and for degree of sulfation by ion exchange chromatography on high pressure liquid chromatography columns. ICF increased the amount of incorporated sulfate per tissue volume unit in the noncalcified epiphyses, but decreased this parameter in the calcified diaphysis. However, in both calcified and noncalcified cartilage, ICF increased the degree of sulfation of the chondroitin sulfate chains. No effects were found on the hydrodynamic size of the PG aggregates or monomers, but in the epiphyses ICF increased the size of the chondroitin sulfate chains. No other changes of structural characteristics of the macromolecules were observed. This study demonstrates that ICF generally stimulated the incorporation of [35S]sulfate into chondroitin sulfate chains. We conclude from the lowered [35S]sulfate content in calcified cartilage that ICF reduced the number of chondroitin sulfate chains and probably PGs while accelerating matrix calcification. It seems likely that the two effects are linked, indicating that a reduction of the number of chondroitin sulfate chains is part of the complicated process of cartilage calcification.


Functional genomics analyses of RNA-binding proteins reveal the splicing regulator SNRPB as an oncogenic candidate in glioblastoma.
Glioblastoma (GBM) is the most common and aggressive type of brain tumor. Currently, GBM has an extremely poor outcome and there is no effective treatment. In this context, genomic and transcriptomic analyses have become important tools to identify new avenues for therapies. RNA-binding proteins (RBPs) are master regulators of co- and post-transcriptional events; however, their role in GBM remains poorly understood. To further our knowledge of novel regulatory pathways that could contribute to gliomagenesis, we have conducted a systematic study of RBPs in GBM. By measuring expression levels of 1542 human RBPs in GBM samples and glioma stem cell samples, we identified 58 consistently upregulated RBPs. Survival analysis revealed that increased expression of 21 RBPs was also associated with a poor prognosis. To assess the functional impact of those RBPs, we modulated their expression in GBM cell lines and performed viability, proliferation, and apoptosis assays. Combined results revealed a prominent oncogenic candidate, SNRPB, which encodes core spliceosome machinery components. To reveal the impact of SNRPB on splicing and gene expression, we performed its knockdown in a GBM cell line followed by RNA sequencing. We found that the affected genes were involved in RNA processing, DNA repair, and chromatin remodeling. Additionally, genes and pathways already associated with gliomagenesis, as well as a set of general cancer genes, also presented with splicing and expression alterations. Our study provides new insights into how RBPs, and specifically SNRPB, regulate gene expression and directly impact GBM development.


Simultaneous backscatter and attenuation estimation using a least squares method with constraints.
Backscatter and attenuation variations are essential contrast mechanisms in ultrasound B-mode imaging. Emerging quantitative ultrasound methods extract and display absolute values of these tissue properties. However, in clinical applications, backscatter and attenuation parameters sometimes are not easily measured because of tissues inhomogeneities above the region-of-interest (ROI). We describe a least squares method (LSM) that fits the echo signal power spectra from a ROI to a three-parameter tissue model that simultaneously yields estimates of attenuation losses and backscatter coefficients. To test the method, tissue-mimicking phantoms with backscatter and attenuation contrast as well as uniform phantoms were scanned with linear array transducers on a Siemens S2000. Attenuation and backscatter coefficients estimated by the LSM were compared with those derived using a reference phantom method (Yao et al. 1990). Results show that the LSM yields effective attenuation coefficients for uniform phantoms comparable to values derived using the reference phantom method. For layered phantoms exhibiting nonuniform backscatter, the LSM resulted in smaller attenuation estimation errors than the reference phantom method. Backscatter coefficients derived using the LSM were in excellent agreement with values obtained from laboratory measurements on test samples and with theory. The LSM is more immune to depth-dependent backscatter changes than commonly used reference phantom methods.


Predicting success in pharmaceutical calculations.
Understanding preadmission predictors of success in pharmacy calculations, an important aspect of pharmacy practice, could impact admissions selection and advising practices. The objective of this study was to determine which student specific preadmission variables best predict success in a pharmaceutical calculations course. Preadmission data from 388 students who completed a one semester pharmaceutical calculations course between 2013 and 2016 were evaluated. This retrospective analysis was performed to determine which of the independent preadmission variables were positively correlated to the pharmaceutical calculations course grade. Fifteen preadmission variables, including demographics, grade point average (GPA), Pharmacy College Admission Test (PCAT) score, prior degrees, and number of pre-professional credit hours completed, were evaluated. Descriptive statistics were used for baseline characteristics and relative importance analysis was used to examine association between the dependent and independent variables. The relative importance analysis revealed eight of the fifteen preadmission variables were significantly correlated with final grades in pharmaceutical calculations. Overall, 26.1% of the variance was explained by these variables. GPA (cumulative and science specific) and PCAT (quantitative and verbal) were the strongest indicators. Preadmission GPA and PCAT scores were the best predictors of success in a pharmaceutical calculations course. About one quarter of the variance was explained by the identified predictive variables, therefore other factors, not evaluated in this study, likely influence the success in pharmaceutical calculations and should be investigated to determine a more dependable set of predictors.


Severity score for hereditary hemorrhagic telangiectasia.
A disease severity score in hereditary hemorrhagic telangiectasia (HHT) would be a useful tool for assessing burden of disease and for designing clinical trials. Here, we propose the first known HHT severity score, the HHT-score. Demographics and disease characteristics were collected for the first 525 HHT patients recruited to the HHT Project of the Brain Vascular Malformation Consortium (BVMC). HHT-score was calculated based on presence of: organ arteriovenous malformations (maximum 3 points); chronic bleeding (maximum 2 points); and severe organ involvement (maximum 2 points). Points were summed and patients categorized as having mild (0-2), moderate (3-4) or severe (5-7) disease. The occurrence of "any adverse outcome" was evaluated for association with HHT-score categories. The frequency of "any adverse outcome" was significantly different across the three groups (49.6% in mild, 65.8% in moderate and 89.5% in severe, p<0.001). Adjusting for age and gender, the risk of "any adverse outcome" was higher in the moderate (OR=1.84, 95% CI: 1.15-2.95, p=0.011) and severe groups (OR=9.16, 95% CI: 1.99-42.09, p=0.004) compared to the mild. We have taken the first steps toward creating a global measure of disease severity in HHT. While the initial results are promising, further validation of the HHT-score is still required.


Insights into the respiratory electron transfer pathway from the structure of nitrate reductase A.
The facultative anaerobe Escherichia coli is able to assemble specific respiratory chains by synthesis of appropriate dehydrogenases and reductases in response to the availability of specific substrates. Under anaerobic conditions in the presence of nitrate, E. coli synthesizes the cytoplasmic membrane-bound quinol-nitrate oxidoreductase (nitrate reductase A; NarGHI), which reduces nitrate to nitrite and forms part of a redox loop generating a proton-motive force. We present here the crystal structure of NarGHI at a resolution of 1.9 A. The NarGHI structure identifies the number, coordination scheme and environment of the redox-active prosthetic groups, a unique coordination of the molybdenum atom, the first structural evidence for the role of an open bicyclic form of the molybdo-bis(molybdopterin guanine dinucleotide) (Mo-bisMGD) cofactor in the catalytic mechanism and a novel fold of the membrane anchor subunit. Our findings provide fundamental molecular details for understanding the mechanism of proton-motive force generation by a redox loop.


Ryanodine receptors contribute to the induction of ischemic tolerance.
Ischemic tolerance (IT) is induced by a variety of insults to the brain (e.g., nonfatal ischemia, heat and hypoxia) and it provides a strong neuroprotective effect. Although the mechanisms are still not fully elucidated, Ca(2+) is regarded as a key mediator of IT. Ryanodine receptors (RyRs) are located in the sarcoplasmic/endoplasmic reticulum membrane and are responsible for the release of Ca(2+) from intracellular stores. In brain, neuronal RyRs are thought to play a role in various neuropathological conditions, including ischemia. The purpose of the present study was to investigate the involvement of RyRs in IT. Pretreatment with a RyR antagonist, dantrolene (25mg/kg, i.p), blocked IT in a gerbil global ischemia model, while a RyR agonist, caffeine (100mg/kg, i.p), stimulated the production of IT. In vitro, using rat hippocampal cells, short-term oxygen/glucose deprivation induced preconditioning and RyR antagonists, dantrolene (50 and 100 μM) and ryanodine (100 and 200 μM) prevented it. RyR protein and mRNA levels were transiently decreased after induction of IT. These results suggest that RyRs are involved in the induction of ischemic tolerance.


Disparities in time trends of cervical cancer mortality rates in Brazil.
This study aimed to correct and describe cervical cancer mortality rates and trends by regions and age-groups in Brazil. It may help planning and implementing policies for cervical cancer control. Data from 2003 to 2012 were accessed through the centralized national mortality database. Correction of the age-specific mortality rates was done by proportional redistribution of ill-defined causes of death and deaths coded as 'uterine, part unspecified'. Annual percentage change (APC) was obtained by trend analysis (Joinpoint regression). In the 10-year period, cancer and ill-defined causes corresponded, respectively, to 18.9 and 10.8 % of all deaths (except injuries). The proportion of ill-defined causes was reduced by more than a half in the period. The age-standardized cervical cancer mortality rate was 7.2 per 100,000 women-years after correction. The total increase in rates after corrections was 50.5 %. A significant decreasing trend in rates was observed at the national level (APC = -0.17, p < 0.001). North was the only region that did not show a decreasing significant trend (APC + 0.07, p = 0.28). Decreasing trends were restricted to age-groups over 40 years. A consistent decreasing trend of cervical cancer mortality rates in Brazil from 2003 to 2012 was observed, although this was not consistent in all regions and restricted to older age-groups. Quality of data needs to be improved. Cancer control policies may consider the differences in access to care and the characteristics of regions to improve their efficiency.


Stochastic prediction in pursuit tracking: an experimental test of adaptive model theory.
In this paper we test the proposition that in pursuit tracking, subjects compute stochastic (statistical) models of the temporal variations in position of the target and use these models to forecast target position for at least a response time interval into the future. A computer simulation of a human operator employing stochastic model prediction of target position is used to generate a synthetic pursuit tracking response signal. Actual pursuit tracking response signals are measured from 10 normal subjects using the same stimulus signal. Cross correlation and spectral analysis are employed to compute gain and phase frequency response characteristics for both synthetic and actual tracking data. The similarity of the gain and phase curves for synthetic and actual data provides compelling evidence in support of the proposition.


Vagotomy during hiatal hernia repair: a benign esophageal lengthening procedure.
This study describes the use of vagotomy in patients during complex laparoscopic esophageal surgery (e.g., reoperative antireflux surgery (rLARS) or paraesophageal hernia (PEH) repair) when, after extensive esophageal mobilization, the gastroesophageal junction cannot be made to reach the abdomen without tension. In doing so, we hope to understand the risk incurred by vagus nerve division in this setting in order to evaluate its role in managing the short esophagus. One hundred and sixty-six patients underwent rLARS or PEH repair between 1/1998 and 6/2003 at our institution. Clinical data was obtained from a prospectively maintained database and systematic patient questionnaires administered for this study. Follow-up was available for 102 (61%) of these patients, at a median of 19 months (range 6-69 months). Fifty-two patients underwent rLARS while 50 patients underwent PEH repair. Thirty patients had a vagotomy during the course of their operation (Vag Group; 20 anterior, six posterior, four bilateral), 13 in the rLARS group (25%), and 17 in the PEH group (34%). The primary presenting symptoms for rLARS and PEH repair patients were improved in 89% in the Vag Group and 91% in the No Vag Group. Similarly, there was no difference in the severity of abdominal pain, bloating, diarrhea, or early satiety between the Vag and No Vag groups at follow-up. No patient required a subsequent operation for gastric outlet obstruction. Vagotomy during rLARS and PEH repair does not lead to a higher rate delayed gastric emptying, dumping syndrome, or other side effects. Thus, we propose vagotomy to be a legitimate alternative to Collis gastroplasty when extensive mobilization of the esophagus fails to provide adequate esophageal length.


Thromboprophylaxis with standard-dose vs. flexible-dose heparin for hospitalized COVID-19 patients: a target trial emulation.
To compare mortality of hospitalized COVID-19 patients under two low-molecular weight heparin (LMWH) thromboprophylaxis strategies: standard dose and variable dose (standard dose increased to intermediate dose in the presence of laboratory abnormalities indicating an increased thrombosis risk). Target trial emulation using observational data from 2,613 adults admitted with a COVID-19 diagnosis in Madrid, Spain between March 16 and April 15, 2020. A total of 1,284 patients were eligible. Among 503 patients without increased baseline thrombotic risk, 28-day mortality risk (95% confidence interval [CI]) was 9.0% (6.6, 11.7) under the standard dose strategy and 5.6% (3.3, 8.3) under the variable dose strategy; risk difference 3.4% (95% CI: -0.24, 6.9); mortality hazard ratio 1.61 (95% CI: 0.97, 2.89). Among 781 patients with increased baseline thrombotic risk, the 28-day mortality risk was 25.8% (22.7, 29.0) under the standard dose strategy and 18.1% (9.3, 28.9) under the intermediate dose strategy; risk difference 7.7% (95% CI: -3.5, 17.2); mortality hazard ratio 1.45 (95% CI: 0.81, 3.17). Major bleeding and LMWH-induced coagulopathy were rare under all strategies. Escalating anticoagulation intensity after signs of thrombosis risk may increase the survival of hospitalized COVID-19 patients. However, effect estimates were imprecise and additional studies are warranted.


Oscillatory chemical reactions: the Tomita-Kitahara model.
Biological clocks share with excitable membrane (nerve, muscle) the requirement that the chemical systems that underlie them have unstable steady states, and hence are capable of limit-cycle oscillations. A particularly simple model of a chemical oscillator is discussed, especially with reference to the reverse reaction, which tends to damp the oscillations. For thermodynamic reasons (entropy production) reversibility can not, however, be neglected. The steady state, even when stable, does not minimize the dissipation function. Computer studies show the response to sinusoidal variation of the rate constant governing the autocatalytic step.


The influence of androstenol - a putative human pheromone - on mood throughout the menstrual cycle.
Each morning for a month female subjects placed either 5 alpha-androst-16-en-3 alpha-ol, a putative human pheromone, or a placebo on the upper lip. Each evening the subjects rated on five scales their moods during that day. In the middle of their monthly cycle those females exposed to androstenol rather than a control tended to rate their moods as submissive rather than aggressive. The compound did not significantly influence ratings of being happy/depressed; lethargic/lively; sexy/unsexy; irritable/good-tempered. The results are discussed in terms of the possible increased olfactory sensitivity of the human female to androstenol in the middle of her monthly cycle.


Remnant lipoproteinemia is a risk factor for endothelial vasomotor dysfunction and coronary artery disease in metabolic syndrome.
This study aimed to determine whether elevated levels of remnant lipoprotein, an atherogenic triglyceride-rich lipoprotein, might be associated with coronary artery disease (CAD) and endothelial vasomotor dysfunction in metabolic syndrome. The fasting serum levels of remnant lipoproteins (remnant-like lipoprotein particles cholesterol; RLP-C) were measured by an immunoseparation method in 210 patients with metabolic syndrome meeting ATP III criteria. Flow-mediated endothelium-dependent dilatation (FMD) in the brachial artery during reactive hyperemia was examined by high-resolution ultrasound technique. This study found that elevated RLP-C levels were a significant and independent risk factor for impaired FMD and angiographically proven coronary artery disease (CAD). Treatment with bezafibrate (n = 20) or atorvastatin (n = 20) for 4 weeks significantly reduced RLP-C levels, with a concomitant improvement in FMD. The % reduction in RLP-C levels from baseline after the treatment was independently correlated with the magnitude of improvement in FMD after adjustment for the % changes in levels of triglyceride, hsCRP, and IL-6, and HOMA index. Thus, elevated levels of RLP-C are a risk factor for CAD and endothelial vasomotor dysfunction, a predictor of coronary events, in metabolic syndrome. Measurement of RLP-C is useful for assessment of CAD risk and therapeutic effects in metabolic syndrome.


Endothelial rehabilitation: the impact of chronic PDE5 inhibitors on erectile function and protein alterations in cavernous tissue of diabetic rats.
Diabetic men generally have reduced efficacy with PDE-5 inhibitors (PDE5i) for the treatment of erectile dysfunction (ED). To determine whether chronic vardenafil exposure alters cavernous protein expression predicting improved erectile function in diabetes. Forty-two adult male Sprague Dawley rats with streptozotocin-induced (50mg/kg IP) diabetes for 4 wk, were exposed to either vehicle or vardenafil for 6 wk. Assessments compared the impact of vardenafil given at 1h and 20 h to erectile function and cellular alterations and downstream translation of cavernous protein profiles were aimed. Vehicle or vardenafil 0.5mg/kg/day by oral gavage for 6 wk. Erectile function, penile tissue morphology, protein expression and surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI) protein profiling were determined. A significant increase of intracavernous pressure was seen in both treatment arms compared to diabetic rats not receiving vardenafil. Immunohistochemical staining showed improved endothelial and smooth muscle cell staining with chronic vardenafil use. Western blot analysis demonstrated increased endothelial cell eNOS and smooth muscle alpha-actin protein content. SELDI protein profiling showed enhanced proteins expression at molecular weights of 14.7, 20, 41.9, 66.2, and 83.9 kDa in the chronically treated vardenafil group. Vardenafil was effective in treating diabetic-induced ED with the greatest effect achieved through chronic dosing, with no additive effect measured with the final acute dose. Changes noted in the histology and protein expression indicate that vardenafil may have a protective effect in this disease state. This finding may serve as a basis for further work evaluating the utility of chronic vardenafil dosing in diabetic men.


Impact of mitral versus aortic bioprosthetic valve position on thromboembolism and bleeding risk in patients with atrial fibrillation.
The impact of valve position on thromboembolic and bleeding events in patients with atrial fibrillation (AF) and bioprosthetic valves is uncertain. We analyzed 159 patients with AF after surgical single-valve replacement from the BPV-AF registry (Retro) (UMIN000034198), which was a multicenter, retrospective, observational registry that enrolled 214 patients with AF and bioprosthetic valves to assess differences in bioprosthetic valve position. Baseline characteristics and clinical outcomes were compared on the basis of the position of aortic or mitral bioprosthetic valves. The primary efficacy endpoint was stroke or systemic embolism, and the primary safety endpoint was major bleeding. There were 85 patients (53.5%) in the aortic valve (AV) group and 74 patients (46.5%) in the mitral valve (MV) group. The MV group had a lower body weight and a higher prevalence of prior major bleeding compared with the AV group. Thromboembolic and bleeding risk scores and the administration of antithrombotic agents were not significantly different between the groups. The primary efficacy endpoint was not significantly different [AV group: 8.2%; 2.25/100 person-years (PY); MV group: 4.1%; 1.01/100 PY] (log-rank, p = 0.23). The primary safety endpoint was significantly higher in the MV group (17.6%; 4.54/100 PY) compared with the AV group (5.9%; 1.59/100 PY) (log-rank, p = 0.049). The adjusted hazard ratio of the primary safety endpoint in the MV group relative to the AV group was 2.71 (95% confidence interval 0.86-8.54, p = 0.09). In Japanese patients with AF and bioprosthetic valves, thromboembolic risk does not differ on the basis of valve position. Bleeding risk is higher in patients with MV bioprostheses, although valve position itself might not be an independent predictor for bleeding.


Should all elderly people receive folate supplements?
Although it was initially thought that folate deficiencies are uncommon in the elderly, the use of the plasma total homocysteine concentration as a metabolic marker of folate status is changing this attitude. Plasma homocysteine measurements in epidemiological studies suggest that subclinical folate deficiencies are common in various populations, including the elderly. The clinical consequences of a compromised folate status may include increased coronary heart disease and cancer risk, and an association between folate deficiency and neuropsychiatric illness has been reported. Although a suboptimal folate status is associated with at least 2 major chronic diseases, it would be premature to recommend folate supplements to all elderly people. We still lack evidence that a suboptimal folate status is causally involved in the pathogenesis of the above mentioned disorders. Furthermore, cancer and the cardiovascular diseases are chronic conditions and it may be too late to use folate supplementation as preventative measures in elderly persons. However, folate supplementation should be considered in elderly people with elevated plasma total homocysteine concentrations and proven cardiovascular disease, in elderly patients treated with drugs known to induce a folate deficiency, and in those who experience neuropsychiatric disorders. Cyanocobalamin (vitamin B12) deficiencies should be excluded before folate supplementation is commenced; if in doubt, it may be safer to supplement folate and vitamin B12 together. The daily folate supplement should be at least 0.5 mg/day and it should never be used as a surrogate for a diet rich in fruit and vegetables.


Attachment styles and psychological profiles of child sex offenders in Ireland.
When 29 child sex offenders, 30 violent offenders, 30 nonviolent offenders, and 30 community controls were compared, a secure adult attachment style was 4 times less common in the child sex offender group than in any of the other three groups. Ninety-three percent of sex offenders had an insecure adult attachment style. Compared with community controls, the child sex offender group reported significantly lower levels of maternal and paternal care and significantly higher levels of maternal and paternal overprotection during their childhood. Compared with all three comparison groups, the child sexual offenders reported significantly more emotional loneliness and a more external locus of control. With respect to anger management, the child sexual offenders' profile more closely approximated those of nonviolent offenders and community controls than that of violent offenders.


[Clinico-physiological parameters of diabetic polyneuropathy in elderly patients with diabetes mellitus type II and the treatment].
Changes of subjective and objective symptoms, elektroneyromiographic parameters, analysis of vegetative system were perfomed in 120 patients with diabetes mellitus type 2. The obtained results show the dependence of subjective and objective manifestations from the duration of the disease.


Effects of low-frequency stimulation on soluble and structure-bound activities of hexokinase and phosphofructokinase in rat fast-twitch muscle.
Several glycolytic enzymes exist in muscle as free and structure-bound forms. A fraction of hexokinase (HK) is associated with the outer mitochondrial membrane. Phosphofructokinase (PFK) and aldolase (ALD) bind to F-actin, and AMP deaminase (AMPase) interacts with myosin. Using low-frequency stimulation (10 Hz, 24 h/d), we studied in rat fast-twitch muscle effects of contractile activity on soluble and structure-bound forms of these enzymes. Phosphoglucose isomerase (PGI), a soluble enzyme, was also examined. Fractional extraction was applied to study the intracellular distribution of soluble and bound enzyme activities 5 min, 1 h, 3 h, 1 d, and 7 d after the onset of stimulation. Confirming previous findings, total HK activity increased 7-fold in 7-d-stimulated muscles, whereas PFK, ALD, and PGI were reduced, ranging between 55% and 80% of their normal activities. AMPase activity was unaltered. At the time points studied, no changes were found in the extraction behavior of PGI and AMPase. The fraction of bound ALD increased slightly (12%). However, the distribution of HK and PFK was markedly altered. Bound PFK increased from 50% in the control to 85% in 7-d-stimulated muscles. Bound HK rose from 52% to 83% during the same time period. The increase in PFK binding was steep and occurred mainly within the first minutes and hours. The increase in HK binding occurred with some delay, but was significant in muscles stimulated for more than 1 h. In view of the altered kinetic properties of F-actin-bound PFK (alleviated allosteric inhibition by ATP) and bound HK (elevated catalytic activity), these changes are interpreted as early responses to match the metabolic demands during maximal contractile activity imposed on a muscle not programmed for sustained activity: Enhanced binding of PFK serves to accelerate glycolytic flux immediately after the onset of stimulation, whereas mitochondrial binding of HK facilitates the phosphorylation of exogenous glucose when glycogen stores have been depleted.


Development of an enhanced formulation for delivering sustained release of buprenorphine hydrochloride.
To control the minimum effective dose, and reduce the number and quantity of administered potent drugs are unique features of advanced drug delivery in situ forming gel formulation. The efficacy, consistency, and increasing the application of existing injection therapies can be enhanced through optimization of controlled released systems by using FDA approved biodegradable PLGA (poly-d,l-lactide-co-glycolide) polymer. The purpose of this study was to develop different in situ forming implant (ISFI) formulations of buprenorphine hydrochloride for post treatment of drug addicts, acute and chronic pains. The drug releases from different ISFIs membranes with and without Tween 80 were compared over a period of time. Kinetic equation followed the Korsmeyer-Peppas model, as the plots showed high linearity. The influence of this additive on polymer properties was investigated using differential scanning calorimetry (DSC), and the membranes structure was studied by X-ray diffractometry (XRD) and scanning electron microscope (SEM). Data revealed that Tween 80 modified the drug release pattern using diffusion mechanism and decreased the glass transition temperature (T g) significantly. The degree of crystallinity was decreased after phase inversion which helps the dissolution of drug from membrane. The porosity of modified membranes was in accordance with release profiles. These findings suggest four different in situ forming implant formulations which can release various dose of the buprenorphine hydrochloride in a prolonged time. Also this surfactant can be an attractive additive for modifying the release rate of drugs from PLGA-based membrane drug delivery systems.


Saudi dental students' perceptions of pediatric behavior guidance techniques.
Dental students receive theoretical and clinical training in pediatric behavioral guidance techniques at university. Therefore, the content of the educational course and the degree of training in behavioral techniques may have an impact on the students' perceptions and practice of such techniques. The purpose of this study was to evaluate Saudi dental students' perceptions of behavior guidance techniques used in pediatric dentistry, and to assess the changes in their perceptions after 1 academic year of a didactic and clinical educational course. This longitudinal study was carried out once at the beginning and once at the end of the 2013/2014 academic year at the College of Dentistry, King Saud University in Riyadh, Saudi Arabia. A questionnaire measuring the perceived acceptability of behavior guidance techniques was completed by 78 fourth-year dental students before and after a pediatric dental course. Acceptability ratings were scored on a 5-point Likert scale and compared and evaluated in relation to demographic data. Paired t-test and one-way analysis of variance were used for the statistical analyses. Before the course, the highest scores were for reinforcement and desensitizing techniques and the lowest were for aversive and communicative techniques. After the course, statistically significant increases were found in the acceptability of aversive techniques (voice control and hand-over-mouth), all pharmacological techniques, and modeling. Most communicative techniques and clinical situations were also rated as significantly more acceptable. Statistically significant decreases in acceptability ratings were found in promising a toy, and immobilization by staff or a parent. Immobilization using a papoose board, modeling, the presence of parents during the child's treatment, and most communicative techniques were rated as significantly more acceptable by male students than female students. In general, Saudi dental students rated most basic behavior guidance techniques as acceptable. An educational course, including didactic and clinical components, improved their acceptability ratings, and had a considerable influence on their perceptions of behavior guidance in pediatric dentistry.


Editorial Commentary: A Steep Learning Curve for Hip Arthroscopy? I Literally Don't Know What This Means Anymore.
Fluoroscopy usage is a common component of hip arthroscopy for femoroacetabular impingement. A surgeon's usage of fluoroscopy (and the subsequent radiation exposure) can decrease over time, as that surgeon gets comfortable with a technique. However, techniques can and usually do change as a surgeon gets increasing experience, and fluoroscopic usage may increase or decrease with different techniques. The term "steep learning curve" should be abandoned, because this term can mean either arduous or easy, depending on context.


Population-based assessment of visual impairment among ethnic Dai adults in a rural community in China.
Dai ethnicity is one of the major Chinese ethnic minorities with a population of about 1.2 million. We aimed to determine the prevalence and potential causes of visual impairment (VI) among ethnic Dai adults aged 50 years or older in a rural community in China. A population-based survey including 2163 ethnic Dai people (80.5%) was undertaken using a random cluster sampling strategy. The detailed eye examination was performed after pupil dilation by trained study ophthalmologists and optometrists. Presenting visual acuity (PVA) and best-corrected visual acuity (BCVA) was measured using the Early Treatment Diabetic Retinopathy Study logMAR chart and VI was defined as a VA of less than 20/63 in the better-seeing eye. The overall prevalence of presenting blindness and low vision was 3.0% (95% CI, 2.3-3.7) and 13.3% (95% CI, 11.9-14.8), respectively. The prevalence estimates were reduced to 2.1% (95% CI, 1.5-2.8) and 6.7% (95% CI, 5.7-7.8) when BCVA was considered. Men were more likely to be affected by low vision but less likely to be blind compared with women. Cataract accounted for 62.7% of presenting low vision and 68.8% of presenting blindness, respectively. In conclusion, VI was a significant health concern in Dai Chinese in China.


Microglial zinc uptake via zinc transporters induces ATP release and the activation of microglia.
Previously, we demonstrated that extracellular zinc plays a key role in transient global ischemia-induced microglial activation through sequential activation of NADPH oxidase and poly(ADP-ribose) polymerase (PARP)-1. However, it remains unclear how zinc causes the sequential activation of microglia. Here, we examined whether transporter-mediated zinc uptake is necessary for microglial activation. Administration of zinc to microglia activated them through reactive oxygen species (ROS) generation and poly(ADP-ribose) (PAR) formation, which were suppressed by intracellular zinc chelation with 25 μM TPEN (N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine) or 2 μM BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester). The (65)Zn uptake by microglia was temperature- and dose-dependent, and it was blocked by metal cations, but not by L-type calcium channel blockers nifedipine and nimodipine. Expression of Zrt-Irt-like protein (ZIP)1, a plasma membrane-type zinc transporter, was detected in microglia, and nickel, a relatively sensitive substrate/inhibitor of ZIP1, showed cis- and trans-inhibitory effects on the (65)Zn uptake. Exposure of microglia to zinc increased the extracellular ATP concentration, which was suppressed by intracellular zinc chelation and inhibition of hemichannels. mRNA expression of several types of P2 receptors was detected in microglia, and periodate-oxidized ATP, a selective P2×7 receptor antagonist, attenuated the zinc-induced microglial activation via NADPH oxidase and PARP-1. Exogenous ATP and 2'(3')-O-(4-benzoyl-benzoyl) ATP also caused microglial activation through ROS generation and PAR formation. These findings demonstrate that ZIP1-mediated uptake of zinc induces ATP release and autocrine/paracrine activation of P2X(7) receptors, and then activates microglia, suggesting that zinc transporter-mediated uptake of zinc is a trigger for microglial activation via the NADPH oxidase and PARP-1 pathway. © 2011 Wiley-Liss, Inc.


Membrane cholesterol plays an important role in enteropathogen adhesion and the activation of innate immunity via flagellin-TLR5 signaling.
Lipid rafts are cholesterol- and sphingolipid-rich ordered microdomains distributed in the plasma membrane that participates in mammalian signal transduction pathways. To determine the role of lipid rafts in mediating interactions between enteropathogens and intestinal epithelial cells, membrane cholesterol was depleted from Caco-2 and IPEC-J2 cells using methyl-β-cyclodextrin. Cholesterol depletion significantly reduced Escherichia coli and Salmonella enteritidis adhesion and invasion into intestinal epithelial cells. Complementation with exogenous cholesterol restored bacterial adhesion to basal levels. We also evaluated the role of lipid rafts in the activation of Toll-like receptor 5 signaling by bacterial flagellin. Depleting membrane cholesterol reduced the ability of purified recombinant E. coli flagellin to activate TLR5 signaling in intestinal cells. These data suggest that both membrane cholesterol and lipid rafts play important roles in enteropathogen adhesion and contribute to the activation of innate immunity via flagellin-TLR5 signaling.


Mitral regurgitation and left atrial thrombus in rheumatic mitral valve disease. A clinicopathologic study.
A total of 255 consecutive patients with rheumatic mitral valve disease, scheduled for surgery, were studied preoperatively by transthoracic echocardiography in a tertiary care hospital setting. Data were analyzed to determine the relationship between mitral regurgitation (MR) and left atrial thrombus (LAT) found at surgery. An LAT was found in 77 patients (30%). Eighty patients had MR of which 30 were mild, 33 were moderate, and 17 showed severe MR. The LAT had an inverse relationship to MR with the prevalence of the former as follows: 37%, 33%, 9%, and 0% in none, mild, moderate, and severe MR, respectively (p < 0.0001). In atrial fibrillation (AF), the prevalence of LAT in patients with predominant MR was 8.3% vs 54% in patients with predominant mitral stenosis (p < 0.0001). From 150 patients with AF, 13 had severe MR and no LAT was found whatsoever. In sinus rhythm, the prevalence of LAT was 0% in predominant MR and 14.3% in patients with predominant mitral stenosis (p < 0.0001). Of 105 patients in sinus rhythm, 14 had moderate or severe MR, none of whom has LAT. When mitral valve area was greater than 1 cm2, the presence of significant MR practically excluded the risk of LAT. In conclusion, rheumatic MR is protective against LAT formation. Prophylactic anticoagulation of symptomatic rheumatic mitral valve disease with AF is not likely to be beneficial when MR is severe or when sinus rhythm is present, and MR is moderate to severe.


A direct enzymatic synthesis of beta-D-galactopyranosyl-D-xylopyranosides and their use to evaluate rat intestinal lactase activity in vivo.
By enzymatic beta-D-galactosylation of D-xylose a mixture of 4-, 3-, and 2-O-beta-D-galactopyranosyl-D-xyloses (1, 4, and 7, respectively) was obtained in 50% isolated yield. Disaccharides 1, 4, and 7 are substrates of intestinal lactase isolated from lamb small intestine with K(m) values of 250.0, 4.5, and 14.0 mM, respectively. The mixture was used to monitor the normal decline in lactase activity in rats that takes place after weaning. The data obtained by this method correlated with the levels of intestinal lactase activity in the same animals after being sacrificed.


Validation of photoplethysmography- and acceleration-based sleep staging in a community sample: comparison with polysomnography and Actiwatch.
Although wrist-worn consumer wearables are widely used for home sleep monitoring, few have been validated. It is unclear whether consumer wearables could be an alternative to Actiwatch. This study aimed to establish and validate an automatic sleep staging system (ASSS) utilizing photoplethysmography and acceleration data collected from a wrist-worn wearable device. Seventy-five participants from a community population underwent overnight polysomnography (PSG) while wearing a smartwatch (MT2511) and Actiwatch Spectrum Plus (Philips Respironics, Inc; Murrysville, PA, USA). Photoplethysmography and acceleration data collected from the smartwatches were utilized to build a 4-stage (wake, light sleep, deep sleep, and rapid eye movement [REM] sleep) classifier, which was validated against PSG. The performance of the sleep/wake classifier was compared with Actiwatch. All analyses were conducted separately for participants with PSG sleep efficiency (SE) ≥ 80% and SE < 80%. The 4-stage classifier and PSG showed fair overall epoch-by-epoch agreement (kappa, 0.55; 95% confidence interval, 0.52 to 0.57). The deep sleep and REM times were comparable between ASSS and PSG, while ASSS underestimated the wake time and overestimated the light sleep time among participants with SE < 80%. Moreover, ASSS underestimated sleep-onset latency and wake after sleep onset and overestimated total sleep time and SE among participants with SE < 80%, while all were comparable among participants with SE ≥ 80%. The biases were smaller for ASSS than for Actiwatch. Our photoplethysmography- and acceleration-based ASSS was reliable for participants with SE ≥ 80% and had a smaller bias than Actiwatch among those with SE < 80%. Thus, ASSS may be a promising alternative to Actiwatch. Registry: ClinicalTrials.gov; Name: Validation of Sleep Healthcare System; URL: https://clinicaltrials.gov/study/NCT04252482; Identifier: NCT04252482. Liu P-K, Ting N, Chiu H-C, et al. Validation of photoplethysmography- and acceleration-based sleep staging in a community sample: comparison with polysomnography and Actiwatch. J Clin Sleep Med. 2023;19(10):1797-1810.


Influence of extracellular pH and perfusion rate on Na+/H+ exchange in cultured opossum kidney cells.
Studies were undertaken in cultured opossum kidney (OK) cells to determine whether the rate of H+ secretion by apical membrane Na+/H+ exchange is modulated by changes in extracellular pH or perfusion rate. H+ secretion was assessed in single cells by measuring the rate of Na(+)-dependent intracellular pH recovery after NH4Cl loading, using the pH-sensitive fluorescent dye, 2'7'-bis(carboxyethyl)-5,6-carboxyfluorescein, in monolayers mounted to allow independent perfusion of the apical and basolateral surfaces. At constant intracellular pH, Na(+)-dependent H+ secretion was found to be inversely related to extracellular H+ activity, and directly related to the perfusate flow rate. Inhibition of H+ secretion by perfusate acidity occurred immediately and was greater when perfusate Na+ was reduced, consistent with H+ competition with Na+ for binding to the transporter. By contrast, the effect of the perfusion rate was a delayed response, requiring 20 min of exposure, and was independent of perfusate Na+ concentration. The results indicate that both extracellular pH and the perfusion rate modulate H+ secretion by OK cells, and that the two effects are independent.


Novel mastoparan analogs induce differential secretion from mast cells.
Cationic amphiphilic peptides stimulate secretion via a receptor-independent action upon G proteins. We have previously utilized chimeric analogs of mastoparan (MP), including galparan (galanin(1-13)-MP ), as molecular probes of secretion. Here, we further resolve the structure-activity relationship of peptidyl secretagogs, including rationally designed chimeric MP analogs. The secretory efficacies of 10 MP analogs were significantly higher than 45 unrelated basic peptides. Comparative studies identified MP analogs that are differential secretagogs for 5-hydroxytryptamine (5-HT) and beta-hexosaminidase. Peptide-induced activation of phospholipase D (PLD), an enzyme intimately involved in regulated exocytosis [5], correlated with the secretion of beta-hexosaminidase but not 5-HT. Thus, these data indicate that different mechanisms are responsible for the exocytosis of 5-HT and beta-hexosaminidase, respectively. Moreover, mastoparan analogs are novel tools for probing the molecular details of exocytosis and other biological phenomena.


Diagnosing and Managing Linear Scleroderma in a Low-Resource Setting.
Linear scleroderma is one of the five forms of scleroderma, but it is the most common form of localized scleroderma in childhood. If left untreated, it can lead to severe disfigurement and functional impairment. The typical appearance is a linear streak with cutaneous induration on the face or head in association with various ophthalmological and neurological signs and symptoms. Treatment typically includes corticosteroids and/or methotrexate with life-long monitoring for recurrence. Case Presentation. A 12-year-old girl presented to our clinic in northern rural Cambodia with a history of a linear streak on her forehead that was growing down her nasal bridge. She denied any tenderness or family history of rheumatic disease. Her history was significant for strabismus as a child. A visiting pediatric rheumatologist assisted us with the appropriate diagnosis and treatment plan. In our case report, we present a child with linear scleroderma who fortunately came to medical attention early and received appropriate treatment before the onset of complications. She was treated with systemic immunosuppression as well as topical steroids. After treatment, she had no further progression on her face and continued to follow up with us to monitor for disease activity. To summarize, linear scleroderma is an uncommon diagnosis for general pediatricians and should be recognized early to provide appropriate treatment and follow-up.


Assessment of the effect of short-term weathering on the molecular-level chemical composition of crude oils in contact with aquatic environments.
Multiple studies have focused on the effect of long-term weathering processes on oils after spill events, without considering the chemical compositional changes occurring shortly after the release of oil into the environment. Therefore, the present study provides a broad chemical characterization for understanding of the changes occurring in the chemical compositions of intermediate (°API = 27.0) and heavy (°API = 20.9) oils from the Sergipe-Alagoas basin submitted to two simulated situations, one under marine conditions and the other in a riverine environment. Samples of the oils were collected during the first 72 h of contact with the simulated environments, followed by evaluation of their chemical compositions. SARA fractionation was used to isolate the resins, which were characterized at the molecular level by UHRMS. The evaporation process was highlighted, with the GC-FID chromatographic profiles showing the disappearance of compounds from n-C10 until n-C16, as well as changes in the weathering indexes and pristane + n-C17/phytane + n-C18 ratios for the crude oils submitted to the riverine conditions. Analysis of the resins fraction showed that basic polar compounds underwent little or no alterations during the early stages of weathering. The marine environment was shown to be much less oxidative than the riverine environment. For both environments, a feature highlighted was an increase of acidic oxygenated compounds with the increase of weathering, especially for the crude oil with °API = 27.0.


Characterization of B16 melanoma-specific cytotoxic T lymphocytes.
A B16 melanoma-specific CD8+ T cell line (AB1) was established from the spleen cells of C57BL/6 mice cured of B16 melanoma with interleukin (IL)-12 treatment. The AB1 line exclusively used T cell receptor Vbeta11. The AB1 cells exhibited a cytolytic activity against both syngeneic B16 melanoma and allogeneic P815 mastocytoma, whereas a cold inhibition assay revealed specificity of the AB1 cells against B16 melanoma. Their lostability to kill a class I loss variant of B16 melanoma was restored by the transfection of H-2Kb gene. In addition, their interferon (IFN)-gamma production was significantly suppressed by the addition of anti-H-2Kb monoclonal antibody, and RT-PCR analysis showed that the AB1 line expressed the mRNA encoding IFN-gamma, but not IL-4 or IL-10. The experiment using synthetic peptides of tyrosinase-related protein-2 (TRP-2) revealed that the AB1 cells could recognize TRP-2(181-188) peptide. Moreover, the AB1 cells showed an in vivo antitumor effect against established pulmonary metastases of B16 melanoma. Overall, these results indicate that the Tc1-type Vbeta11+ AB1 cells exert an antitumor activity against syngeneic B16 melanoma through recognition of TRP-2(181-188) peptide in an H-2Kb-restricted manner.


Biliary Epithelial Senescence in Cellular Rejection Following Live Donor Liver Transplantation.
As with the hepatocytes, cholangiocyte senescence can also easily be detected in damaged small bile ducts and bile ductules during liver disease affecting the biliary system and cholangiocytes. Despite cellular senescence being a feature of chronic progressive cholangiopathies in adults, only a few studies have investigated its role in liver transplant rejection. Transplant biopsies displaying features of rejection were reviewed and classified based on the type of rejection and the time since transplantation. An immunohistochemistry panel has been applied for 3 senescent cell markers (p53, p21, p16). Immunohistochemical expression analysis for the biliary senescence markers (53 biopsies) was done in the post-transplantation periods (Group 1-4) for the cases with the histologically proven diagnosis of rejection. In post-transplant group 1 (<3 months), group 2 (3-6 months), group 3 (6-12 months) and group 4 (>12 months), any 2 senescent markers' positivity was noted in 5/14 (35.7%), 8/13 (61.5%), 16/17 (94.1%) and 9/9 (100%) biopsies respectively and were comparable in all four groups (P = 0.001). A comparison of early biopsies (Group1; 3 months) and late biopsies (Group 2,3&4; >3 months) revealed significantly higher expression in late biopsies (>3 months) (P = 0.001 for any two markers). In ACR, LAR, ECR, and CR/DR any two senescent markers were positive in 14/28 (50%), 12/13 (92.3%) cases, 9/9 (100%), and 3/3cases (100%). Senescent markers (any two) were comparable in all four histological groups (P < 0.001).LAR group had increased expression (P = 0.009 for any two markers and 0.001 for all three markers) and has increased progression to CR (P = 0.019) as compared to ACR. This study on a large number of LDLT allograft biopsies demonstrates the role of biliary senescence in rejection and suggests a pathobiological role for senescence in the poor prognosis seen in late acute cellular rejection and chronic rejection.


Investigation of correlations between chemical parameters of metal ions and acute toxicity in mice and Drosophila.
We are studying correlations between physicochemical properties associated with metal ions and observed toxicity. In order to test correlations, we obtained, under uniform conditions, LD50 values for acute toxicity in mice for 24 metal ions. The new data show a better correlation between LD50 and Pearson and Mawby's softness parameter sigma p, defined using chemical concepts of hard and soft acids and bases, than had been obtained by others. From a wide range of physicochemical parameters, the electrode potential can give almost as good a correlation as sigma p. Better correlations might exist for parameters more relevant to biological systems.


[Presence, in ticks from the Geneva region, of infective larvae which may be related to the dog filaria Dipetalonema grassii].
About forty infective filarial larvae were collected from Rhipicephalus sanguineus (two ticks positive out of about 50 examined) living inside a house and presumably originally transported from the south of France or Toscana. The larvae are of the genus Dipetalonema; they are distinct from infective stages of the three most common European species, D. dracunculoides and D. reconditum from dogs and D. rugosicauda from roe deer. They may however be larvae of D. arassii (Noé, 1907), a dog filarid which has not been reported since its original descovery in Italy.


Electronmicroscopic investigation of the flexibility of F-actin.
The contour lenghts and the end-to-end distances of a large number of F-actin filaments were measured in electronmicrographs. Preparation of F-actin for electron-microscopy was made at three different temperatures. The flexibility parameter or the elastic modulus for bending of F-actin was determined from the relation between the contour length and the end-to-end distance after statistical treatment. The value of the flexibility parameter obtained here showed good agreement with that obtained by the quasielastic light scattering at all temperatures examined. The flexibility of F-actin increased with raising temperature and with the binding of heavy meromyosin.


Crystalloid strong ion difference determines metabolic acid-base change during acute normovolaemic haemodilution.
To study the acid-base effects of crystalloid strong ion difference (SID) during haemodilution. Prospective in vivo study. University laboratory. Anaesthetised, mechanically ventilated Sprague-Dawley rats. Rats were studied in seven groups of three. Each group underwent normovolaemic haemodilution with one of seven crystalloids, with SID values from 0 to 40 mEq/l. Six exchanges of 9 ml crystalloid for 3 ml blood were performed. [Hb] fell from 142+/-17 to 44+/-10 g/l ( p<0.0001). Final plasma [lactate] was 1.1+/-0.6 mmol/l. Final standard base excess values ranged from -8 to +7 mmol/l, and were directly correlated with crystalloid SID (R(2)=0.91). By linear regression, the SID of a crystalloid balanced to maintain standard base excess=0 mmol/l was 23.7 mEq/l. Dilutions 2-5 produced similar results. There is a linear relationship between crystalloid SID and post-dilutional metabolic acid-base status. The SID of a crystalloid balanced for normovolaemic haemodilution is 24 mEq/l. These principles are applicable in designing fluids for volume resuscitation, acute normovolaemic haemodilution and cardio-pulmonary bypass.


Diverse redox-active molecules bearing identical thiol-terminated tripodal tethers for studies of molecular information storage.
To examine the effects of molecular structure on charge storage in self-assembled monolayers (SAMs), a family of redox-active molecules has been prepared wherein each molecule bears a tether composed of a tripodal linker with three protected thiol groups for surface attachment. The redox-active molecules include ferrocene, zinc porphyrin, ferrocene-zinc porphyrin, magnesium phthalocyanine, and triple-decker lanthanide sandwich coordination compounds. The tripodal tether is based on a tris[4-(S-acetylthiomethyl)phenyl]-derivatized methane. Each redox-active unit is linked to the methane vertex by a 4,4'-diphenylethyne unit. The electrochemical characteristics of each compound were examined in solution and in SAMs on Au. Redox-kinetic measurements were also performed on the SAMs (with the exception of the magnesium phthalocyanine) to probe (1) the rate of electron transfer in the presence of an applied potential and (2) the rate of charge dissipation after the applied potential is disconnected. The electrochemical studies of the SAMs indicate that the tripodal tether provides a more robust anchor to the Au surface than does a tether with a single site of attachment. However, the electron-transfer and charge-dissipation characteristics of the two tethers are generally similar. These results suggest that the tripodal tether offers superior stability characteristics without sacrificing electrochemical performance.


Treatment of sternal wound infections after open-heart surgery.
The aim of this study was to investigate the proper treatment of infected median sternotomy wounds. A retrospective study was conducted to investigate the proper treatment of infected median sternotomy wounds on patients with sternal wound infections from January 2007 to July 2009. The characteristics of the sternal infections and the treatment outcomes were analysed. Ninety-seven patients with sternal wound infections were treated. A total of 32 patients acquired the infection within one month after open-heart surgery, 10 patients got the infection one to two months after the surgery, and 1 patient died two days after debridement. There were 54 patients who acquired the infection beyond two months post-surgery, while 1 patient died on the day before the operation. One patient received four cycles of wound debridement, 18 patients received two cycles and 78 patients only received one cycle. A total of 14 patients received a vacuum-assisted closure treatment. There were 73 patients who had surgery for repair of muscle flaps, 1 patient for breast tissue flap, 63 patients for pectoralis major muscle flap, and 9 patients with rectus abdominis muscle flap. There were 12 patients who received a transverse plate fixation of the sternum with titanium plating. A positive prognosis can be obtained by the algorithm treatment based on the onset and depth of the sternal infection.


Study regarding the survival of patients suffering a traumatic cardiac arrest.
Severe trauma is the most frequent cause of death in young people, in civilized countries with major social and vital costs. The speed of diagnostic decision making and the precocity of treatment approaches are both essential and depend on the specialists' colaboration. The present study aims to emphasize the actual situation of medical interventions in case of cardiorespiratory arrest due to trauma. 1387 patients who suffered a cardio respiratory arrest both traumatic and non-traumatic were included in order to point out the place of traumatic arrest. Resuscitation of such patients is considered useless and resource consumer by many trauma practitioners who are reporting survival rates of 0%-3.5%. As the determinant of lesions, trauma etiology was as it follows car accidents - 43%, high falls - 30%, suicidal attempts - 3%, domestic violence - 3%, other causes - 21%. Hypovolemia remains the major cause of cardiac arrest and death and that is why the efforts of emergency providers (trauma team) must be oriented towards "hidden death" in order to avoid it. This condition could be revealed and solved easier with minimal diagnostic and therapeutic maneuvers in the emergency department.


Small bowel permeability to 51Cr-EDTA in children with recurrent abdominal pain.
Small bowel permeability was investigated in 87 children with recurrent abdominal pain by measuring the 24-h urinary excretion of orally administered 51Cr-EDTA. The mean excretion was 3.64% +/- 1.49% per 24 h. The difference between the mean urinary excretion in children with recurrent abdominal pain and control children (2.51% +/- 0.70%), was significant (p less than 0.01, two sample t-test). The increased small bowel permeability in children with recurrent abdominal pain might indicate an intestinal etiology for the patients' complaints.


Antigen-induced neutrophil chemotactic activity from sensitized lung.
We have used the model of ovalbumin-sensitized guinea pigs to define the source of antigen-derived, high-molecular-weight neutrophil chemotactic activity (NCA). Incubating sensitized lung fragments with specific antigen (ovalbumin) but not irrelevant antigen (bovine serum albumin) or buffer resulted in the release of high-molecular-weight NCA and histamine, suggesting a lung mast cell origin for NCA. High-molecular-weight NCA accounted for greater than 90% of the NCA observed after antigen-lung incubation. The importance of high-molecular-weight NCA in recruitment of neutrophilic leukocytes to the site of allergic inflammatory reactions is emphasized.


Facile air-oxidation of large aromatic hydrocarbon bay regions to bay region quinones: predicted oxygen-sensitivity of hydrogen-terminated carbon nanotubes.
Dimesitylbisanthene (1) oxidizes to the corresponding bay region quinone (2) on standing in solutions exposed to air and ambient light. It is anticipated that hydrogen-terminated carbon nanotubes with bay regions on the rim are likely to exhibit even greater sensitivity toward air oxidation.


Local angiotensin II contributes to tumor resistance to checkpoint immunotherapy.
Current checkpoint immunotherapy has shown potential to control cancer by restoring or activating the immune system. Nevertheless, multiple mechanisms are involved in immunotherapy resistance which limits the clinical benefit of checkpoint inhibitors. An immunosuppressive microenvironment is an important factor mediating the original resistance of tumors to immunotherapy. A previous report by our group has demonstrated that local angiotensin II (AngII) predominantly exists in a tumor hypoxic microenvironment where hypoxic tumour cells produced AngII by a hypoxia-lactate-chymase-dependent mechanism. Here, using 4T1 and CT26 syngeneic mouse tumor models, we found that local AngII in the tumor microenvironment was involved in immune escape of tumour cells and an AngII signaling blockage sensitized tumours to checkpoint immunotherapy. Furthermore, an AngII signaling blockage reversed the tumor immunosuppressive microenvironment, and inhibition of angiotensinogen (AGT, a precursor of AngII) expression strongly triggered an immune-activating cytokine profile in hypoxic mouse cancer cells. More importantly, AGT silencing combined with a checkpoint blockage generated an abscopal effect in resistant tumors. Our study demonstrated an important role of local AngII in the formation of a tumor immunosuppressive microenvironment and its blockage may enhance tumor sensitivity to checkpoint immunotherapy. The combination of an AngII signaling blocker and an immune-checkpoint blockage could be a promising strategy to improve tumors responses to current checkpoint immunotherapy.


Mastocytosis.
Mastocytosis is defined by a pathological increase in mast cell numbers in tissues. Recent clinical observations on rare manifestations highlight the diversity of this disease. The diagnosis is now aided by new surrogate markers. At the molecular level, recent studies have reinforced the role of activating mutations in KIT in the etiology of mastocytosis. These findings provide a conceptual basis for the development for new therapeutic strategies.


Long-term adverse effects on dentition in children with poor-risk neuroblastoma treated with high-dose chemotherapy and autologous stem cell transplantation with or without total body irradiation.
Chemo- and radiotherapy may have injurious effects on developing teeth. In this long-term follow-up study among poor-risk neuroblastoma (NBL) survivors our aims were: (1) to assess both the type and extent of the side-effects of the anticancer treatment on tooth development; and (2) to develop an index for expressing total damage to the permanent dentition. We studied the dental development from panoramic radiographs (PRG) of 18 long-term survivors treated under the age of 6 years with high-dose (HD) chemotherapy and autologous stem cell transplantation (ASCT) for poor-risk NBL. The myeloablative therapy was either HD chemotherapy and fractionated total body irradiation (TBI) of 10-12 Gy (TBI group, n = 10) or HD chemotherapy only (non-TBI group, n = 8). A defect index (DeI) was developed to describe the damage to the permanent dentition. The DeI was also tested in 18 healthy adolescents. All NBL patients had disturbances in dental development including short roots, arrested root development, microdontia and tooth aplasia. After TBI, 9/10 patients had very severe root defects, in contrast to none in the non-TBI group. All children in the TBI group had 2-12 (mean 6.6) missing permanent teeth, while 2/5 in the non-TBI group (3/8 excluded due to young age) had two and four missing permanent teeth, respectively. Microdontia was found at equal frequency in both groups. The mean value of the DeI was 70.0 (range 28-117) in the TBI group, 15.2 (range 4-34) in the non-TBI group (P<0.001, Mann-Whitney U test) and 1.8 (range 0-15) in healthy adolescents. Disturbances in dental development may compromise occlusal function in poor-risk NBL patients after ASCT, especially when TBI is included in the conditioning regimen. Long-term dental follow-up and rehabilitation is required.


Automated peritoneal dialysis-related peritonitis due to Salmonella enteritidis in a pediatric patient.
Peritoneal dialysis (PD) represents the preferred renal replacement therapy in pediatric patients; however, the younger the age at PD initiation, the greater the risk of PD-related infections. We present here the first case report of a 14-year-old girl with automated peritoneal dialysis-related Salmonella enteritidis peritonitis. The child responded only partially to an appropriate course of antibiotic therapy; the PD catheter was therefore removed and the patient shifted to hemodialysis. Along with the clinical course description, we review the mechanisms by which this group of Salmonella could infect the peritoneal cavity in patients on PD. Pediatric nephrologists should be aware that peritoneal catheter removal is often required for the complete resolution of this type of bacterial peritonitis.


Patterns of parasitism in insular lizards: effects of body size, condition and resource availability.
The spatial distribution of a population of lizards is influenced by the distribution of resources and the individuals' skills to gain access to them. On several Mediterranean islands with food scarcity and unpredictable resource availability, some resources can be extremely important during some periods of the year, as, e.g., the dead horse arum, Dracunculus muscivorus, on Aire Island, Spain. In this study, we analysed parasite infestation in an endemic lizard, Podarcis lilfordi, in relation to the distribution of D. muscivorus during spring. We found a higher prevalence of parasitism in males from a central area where the plant was significantly more abundant. Also, in this area, males were more abundant and larger than in two areas with lower plant densities. Moreover, overall differences appeared in the intensity of infection, which was higher in individuals from the central area, and we found a relationship between body size and parasite load, with more infected cells in larger individuals. Spatial distribution of individuals due to differential availabilities of D. muscivorus is proposed as an explanation for the observed differences.


Learning from the past in the COVID-19 era: rediscovery of quarantine, previous pandemics, origin of hospitals and national healthcare systems, and ethics in medicine.
After the dramatic coronavirus outbreak at the end of 2019 in Wuhan, Hubei province, China, on 11 March 2020, a pandemic was declared by the WHO. Most countries worldwide imposed a quarantine or lockdown to their citizens, in an attempt to prevent uncontrolled infection from spreading. Historically, quarantine is the 40-day period of forced isolation to prevent the spread of an infectious disease. In this educational paper, a historical overview from the sacred temples of ancient Greece-the cradle of medicine-to modern hospitals, along with the conceive of healthcare systems, is provided. A few foods for thought as to the conflict between ethics in medicine and shortage of personnel and financial resources in the coronavirus disease 2019 era are offered as well.


The non-human primate oocyte and embryo as a model for women, or is it vice versa?
The role of the non-human primate (NHP) oocyte and embryo in translational research is considered here including both in vitro activities directly involving oocytes or embryos as well as animal studies that impact reproductive function. Reasons to consider NHPs as animal research models along with their limitations are summarized. A case is made that in limited instances, such as in the development and application of the assisted reproductive technologies or in the study of embryonic stem cells, the human oocyte and embryo have acted as models for the monkey. The development of strategies for the preservation of fertility is used as an example of ongoing research in the non-human primate that cannot be conducted in women for ethical reasons. In animal studies, monitoring reproductive potential, responses to embryonic stem cell transplantation, along with translational research in the field of contraceptive development for women are considered as subjects that benefit from the availability of a NHP model.


Diazo dye Congo Red degradation using a Boron-doped diamond anode: An experimental study on the effect of supporting electrolytes.
Diazo dye Congo Red (CR) solutions at 100mg/L, were degraded using different supporting electrolytes in an electrochemical advanced oxidation process (EAOPs), like the anodic oxidation (AOx/BDD). All experiments were carried out in a 3L flow reactor with a Boron-doped diamond (BDD) anode and stainless steel cathode (AISI 304), at 7.5, 15, 30 and 50mA/cm(2) current densities (j). Furthermore, each experiment was carried out under a flow rate of 7L/min. Additionally, HClO4, NaCl, Na2SO4, and H2SO4 were tested as supporting electrolytes at a 50mM concentration. The degradation process was at all times considerably faster in NaCl medium. Solutions containing SO4(2-) or ClO4(-) ions were less prompted to degradation due to the low oxidation power of these species into the bulk. Dissolved organic carbon (DOC) analysis, was carried out to evaluate the mineralization of CR. The degradation of CR, was evaluated with the HPLC analysis of the treated solutions.


Determination of benzene, toluene, ethylbenzene and xylenes in indoor air at environmental levels using diffusive samplers in combination with headspace solid-phase microextraction and high-resolution gas chromatography-flame ionization detection.
An improved analytical method for passive air sampling is presented based on a combination of commercially available diffusive samplers with headspace solid-phase microextraction and high-resolution gas chromatography with flame ionization detection (HRGC-FID). This procedure is targeted for short-term BTEX (benzene, toluene, ethylbenzene and o-, m- and p-xylenes) determinations at environmental concentrations and can be applied for sampling intervals between 30 min and 24 h. The analytes are adsorbed onto the charcoal pad of a passive sampler and then extracted with carbon disulphide-methanol. After removal of the carbon disulphide by xanthation, the BTEXs are enriched on a Carboxen SPME fiber, thermally desorbed and analysed by HRGC-FID. Detection limits for a sampling interval of 2 h are between 0.4 and 2 micrograms/m3, within-series precision ranges between 6.6 and 12.8%, day-to-day precision is between 11.1 and 15.2%. The results obtained with this procedure are validated by comparison with active sampling. Detection limits and a further reduction of the sampling time are limited by blanks of the chemicals and the diffusive samplers. Procedures to eliminate these blanks are described in detail. Applications such as the determination of BTEXs in indoor air inside buildings, inside a train and a car are presented, indicating the usefulness of the described procedure for short-term measurements of environmental BTEX concentrations. An advantage of passive samplers is the storage stability for at least six months, which is essential for its use in large epidemiological studies.


Is roxithromycin better than amoxicillin in the treatment of acute lower respiratory tract infections in primary care? A double-blind randomized controlled trial.
To assess the efficacy of roxithromycin relative to amoxicillin. We conducted a double-blind randomized controlled trial of oral 500 mg amoxicillin 3 times per day vs oral 300 mg roxithromycin once a day for 10 days. We included 196 adults who had presented to a general practitioner with lower respiratory tract infection (LRTI) and, in the physician's opinion, needed antibiotic treatment. We measured clinical response after 10 and 28 days, defined in 4 ways: (1) decrease in LRTI symptoms; (2) complete absence of symptoms; (3) decrease in signs; and (4) complete absence of signs. Self-reported response included the decrease in symptoms and the time until resumption of impaired or abandoned daily activities on days 1 through 10, 21, and 27. Clinical cure rates after the completion of antibiotic treatment (10 days) were not significantly different for the 2 groups. After 28 days, the roxithromycin group showed no increase in cure rate as evidenced by the decrease in symptoms, indicating a significantly lower cure rate. However, this difference did not alter physicians' overall conclusion after complete follow-up that 90% of patients, regardless of age, had been effectively treated with either amoxicillin or roxithromycin. The surplus value of roxithromycin was not confirmed. Amoxicillin remains a reliable first-choice antibiotic in the treatment of LRTI in general practice.


Development of a New Murine Model of Type 2 Autoimmune Hepatitis Using a Human Liver Protein.
Autoimmune hepatitis (AIH) is a chronic inflammatory condition of the liver characterized by parenchymal destruction, hypergammaglobulinemia, specific autoantibody production, and hepatic fibrosis and necrosis. Murine models of AIH have been described; however, little is known about the immunologic mechanisms of tissue destruction. In this study, a new murine model of type 2 AIH was developed using recombinant human cytochrome P450 (CYP) 2D6 emulsified with complete Freund's adjuvant (CFA). BALB/c mice were immunized with 2 μg/mL i.p. of CYP2D6 in CFA. The control group received CFA or phosphate-buffered saline alone. Alanine aminotransferase activity, autoantibody production, IgG concentrations, histologic damage, and specific T-cell response were evaluated. Persistent AIH, characterized by cellular infiltration, hepatic fibrosis, elevated alanine aminotransferase, and the production of anti-liver kidney microsomal antibody type 1 developed in CFA/CYP2D6-immunized mice. These mice presented high levels of IgG and its subclasses IgG1, IgG2a, and IgG2b against liver self-proteins. Interestingly, IL-2+ and interferon γ-positive Cyp2d6-specific T cells were present in greater concentrations in mice immunized with CFA/CYP2D6 compared with control. Immunization with CFA, in combination with a natural human autoantigen like CYP2D6, was demonstrated to break tolerance, resulting in a chronic form of autoimmune-related liver damage. This murine model of type 2 AIH is expected to be instrumental in understanding the immunologic mechanisms of the pathogenesis of this autoimmune liver disease.


Strontium (Sr) and silver (Ag) loaded nanotubular structures with combined osteoinductive and antimicrobial activities.
Two frequent problems are associated with the titanium surfaces of bone/dental implants: lack of native tissue integration and associated infection. These problems have prompted a significant body of research regarding the modification of these surfaces. The present study describes a hydrothermal treatment for the fabrication of strontium (Sr) and silver (Ag) loaded nanotubular structures with different tube diameters on titanium surfaces. The Sr loading from a Sr(OH)2 solution was regulated by the size of the inner diameter of the titanium nanotubes (NT) (30nm or 80nm, formed at 10V or 40V, respectively). The quantity of Ag was adjusted by immersing the samples in 1.5 or 2.0M AgNO3 solutions. Sr and Ag were released in a controllable and prolonged matter from the NT-Ag.Sr samples, with negligible cytotoxicity. Prominent antibacterial activity was observed due to the release of Ag. Sr incorporation enhanced the initial cell adhesion, migration, and proliferation of preosteoblast MC3T3-E1 cells. Sr release also up-regulated the expression of osteogenic genes and induced mineralization, as suggested by the presence of more mineralized calcium nodules in cells cultured on NT-Ag.Sr surfaces. In vivo experiments showed that the Sr-loaded samples accelerated the formation of new bone in both osteoporosis and bone defect models, as confirmed by X-ray, Micro-CT evaluation, and histomorphometric analysis of rats implanted with NT-Ag.Sr samples. The antibacterial activity and outstanding osteogenic properties of NT-Ag.Sr samples highlight their excellent potential for use in clinical applications. Two frequent problems associated with Ti surfaces, widely used in orthopedic and dental arenas, are their lack of native tissue integration and risk of infection. We describe a novel approach for the fabrication of strontium (Sr) and silver (Ag) loaded nanotubular structures on titanium surfaces. A relevant aspect of this work is the demonstration of long-lasting and controllable Ag release, leading to excellent antibacterial and anti-adherent properties against methicillin-resistant Staphylococcus aureus (MRSA), and Gram-negative bacteria such as Escherichia coli. The extended release of Sr accelerates the filling of bone defects by improving the repair of damaged cortical bone and increasing trabecular bone microarchitecture. Our results highlight the potential of Sr and Ag loaded nanotubular structures for use in clinical applications.


The evolution of contact prediction: evidence that contact selection in statistical contact prediction is changing.
Over the last few years, the field of protein structure prediction has been transformed by increasingly accurate contact prediction software. These methods are based on the detection of coevolutionary relationships between residues from multiple sequence alignments (MSAs). However, despite speculation, there is little evidence of a link between contact prediction and the physico-chemical interactions which drive amino-acid coevolution. Furthermore, existing protocols predict only a fraction of all protein contacts and it is not clear why some contacts are favoured over others. Using a dataset of 863 protein domains, we assessed the physico-chemical interactions of contacts predicted by CCMpred, MetaPSICOV and DNCON2, as examples of direct coupling analysis, meta-prediction and deep learning. We considered correctly predicted contacts and compared their properties against the protein contacts that were not predicted. Predicted contacts tend to form more bonds than non-predicted contacts, which suggests these contacts may be more important than contacts that were not predicted. Comparing the contacts predicted by each method, we found that metaPSICOV and DNCON2 favour accuracy, whereas CCMPred detects contacts with more bonds. This suggests that the push for higher accuracy may lead to a loss of physico-chemically important contacts. These results underscore the connection between protein physico-chemistry and the coevolutionary couplings that can be derived from MSAs. This relationship is likely to be relevant to protein structure prediction and functional analysis of protein structure and may be key to understanding their utility for different problems in structural biology. We use publicly available databases. Our code is available for download at https://opig.stats.ox.ac.uk/. Supplementary information is available at Bioinformatics online.


Role of a nonsteroidal anti-inflammatory agent, ibuprofen, in coronary revascularization after acute myocardial infarction.
The efficacy of using a nonsteroidal anti-inflammatory agent such as ibuprofen for the salvage of ischemic and reperfused myocardium was investigated by examining its ability to improve global and regional functions as well as to preserve high-energy phosphate compounds and inhibit creatine kinase release from an isolated in-situ pig heart subjected to 1 h of normothermic regional ischemia followed by 1 h of global hypothermic arrest and 1 h of normothermic reperfusion. Preperfusion of the heart for 15 min prior to ischemic insult with 50 microM ibuprofen failed to mitigate the myocardial reperfusion injury. Ibuprofen, however, functioned as an anti-inflammatory agent, as judged by its ability to inhibit the influx of indium-111-labeled polymorphonuclear leukocytes and chromium-51 (51Cr)-labeled platelets into the ischemic and reperfused heart. It also blocked the cyclooxygenase pathway, as evidenced by the significant reduction of 6-keto-prostaglandin F1 alpha and thromboxane B2 concentrations in the perfusate. Inhibition of cyclooxygenase resulted in increased accumulation of nonesterified fatty acids, particularly arachidonic acid, in the heart. These results suggest that although ibuprofen can inhibit polymorphonuclear leukocyte and platelet influx into the ischemic and reperfused heart, it causes further damage to the already ischemic heart by reducing prostacyclin concentration and increasing free fatty acids in the heart.


Prognostic impact of pathological response to neoadjuvant chemotherapy followed by definitive surgery in sinonasal squamous cell carcinoma.
The significance of neoadjuvant chemotherapy followed by definitive surgery for sinonasal squamous cell carcinoma (SCC) was investigated using surgical specimens. Surgery was performed in 58 patients, including 43 patients who had received neoadjuvant chemotherapy. The pathological response was classified as grades 0 (no effect), 1 (slight effect), 2 (moderate effect), and 3 (marked effect), and its correlation with prognosis was investigated. Grade 2 or 3 response was observed in 34.9% of cases that received neoadjuvant chemotherapy. Overall survival (OS), disease-free survival (DFS), locoregional control, and freedom from distant metastasis were significantly better in the effective neoadjuvant chemotherapy group (grades 2 and 3) than in the less effective neoadjuvant chemotherapy group (grades 0 and 1) and the non-neoadjuvant chemotherapy group combined. Pathological response to neoadjuvant chemotherapy in sinonasal SCC was related to disease prognosis. © 2016 Wiley Periodicals, Inc. Head Neck 38: E1305-E1311, 2016.


Incidence and Mortality Risks of Cancer in Patients with Type 2 Diabetes: A Retrospective Study in Shanghai, China.
Evidence from epidemiologic investigation indicates that people with type 2 diabetes (T2DM) are at a significantly higher risk of many types of cancer and mortality. The aim of this study was to investigate the incidence and mortality risks of cancer in patients with T2DM compared with the general population in Shanghai, China. Based on the Shanghai Diabetes Registry (SDR) database linking to the Shanghai Cancer Registry and Surveillance System (SCRSS), a total of 12,276 T2DM patients without cancer were defined and followed up from 1 December 2001 to 31 July 2011. Standardized incidence ratio (SIR) and standardized mortality ratio (SMR) with 95% confidence interval (CI) were calculated using the whole gender and age-matched general population of Shanghai as a reference during the same period. The overall cancer risk was found higher in both males and females T2DM patients, with the SIR of 3.14 (95% CI 2.73-3.56) and 4.29 (95% CI 3.64-4.94), respectively. The overall mortality risk of cancer also significantly increased with the SMR of 2.27 (95% CI 1.86-2.68) and 1.86 (95% CI 1.46-2.26), respectively. Pancreatic cancer was with the highest SIR and SMR in both genders. Compared with the general population, patients with T2DM were associated with higher incidence and mortality risks of cancer, especially pancreatic cancer.