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ABSTRACTNociceptor sensitization following nerve injury or inflammation leads to chronic pain. An increase in the nociceptor hyperpolarization-activated current, Ih, is observed in many models of pathological pain. Pharmacological blockade of Ihprevents the mechanical and thermal hypersensitivity that occurs during pathological pain. Alterations in the Hyperpolarization-activated Cyclic Nucleotide-gated ion channel 2 (HCN2) mediate Ih-dependent thermal and mechanical hyperalgesia. Limited knowledge exists regarding the nature of these changes during chronic inflammatory pain. Modifications in HCN2 expression and post-translational SUMOylation have been observed in the Complete Freund’s Adjuvant (CFA) model of chronic inflammatory pain. Intra-plantar injection of CFA into the rat hindpaw induces unilateral hyperalgesia that is sustained for up to 14 days following injection. The hindpaw is innervated by primary afferents in lumbar DRG, L4-6. Adjustments in HCN2 expression and SUMOylation have been well-documented for L5 DRG during the first 7 days of CFA-induced inflammation. Here, we examine bilateral L4 and L6 DRG at day 1 and day 3 post-CFA. Using L4 and L6 DRG cryosections, HCN2 expression and SUMOylation were measured with immunohistochemistry and proximity ligation assays, respectively. Our findings indicate that intra-plantar injection of CFA elicited a bilateral increase in HCN2 expression in L4 and L6 DRG at day 1, but not day 3, and enhanced HCN2 SUMOylation in ipsilateral L6 DRG at day 1 and day 3. Changes in HCN2 expression and SUMOylation were transient over this time course. Our study suggests that HCN2 is regulated by multiple mechanisms during CFA-induced inflammation.

Background and Aims: The aim of this study was to evaluate the effect of smoking and body mass index on the occurrence of complications after alloplastic breast reconstruction.Materials and Methods: A consecutive series of 56 patients treated with immediate or delayed alloplastic breast reconstruction, including six cases combined with latissimus dorsi flap, at three hospitals between 2012 and 2018 were included. Complications were scored and defined according to Clavien–Dindo. To evaluate the impact of smoking, body mass index, and other potential risk factors on the occurrence of any and severe complications, univariate and multivariate logistic regression analyses were applied to estimate odds ratios and 95% confidence intervals.Results: In 56 patients, 22 patients had a complication. As much as 46% of smokers had severe complications compared to 18% of non-smokers. Of patients with body mass index ⩾ 25, 40% had severe complications compared to 10% with body mass index < 25. Smokers had eight times more chance of developing severe complications than non-smokers (ORadjusted= 8.0, p= 0.02). Patients with body mass index ⩾ 25 had almost 10 times more severe complications compared to patients with body mass index ⩽ 25 (ORadjusted= 9.9, p= 0.009). No other risk factors were significant.Conclusion: Smoking and body mass index ⩾ 25 both increased the complication rate to such an extent that patients should be informed about their increased risk for complications following alloplastic breast reconstruction and on these grounds surgeons may delay alloplastic breast reconstruction. It is an ethical dilemma whether one should deny overweight and obese patients and those who smoke an immediate alloplastic breast reconstruction. For both life style interventions, adequate guidance should be made available.

BACKGROUND: The tendency of sepsis to progress rapidly and the benefit of an early start of treatment emphasise the importance of fast risk stratification in the emergency department (ED). The aim of the present work was to validate the Mortality in Emergency Department Sepsis (MEDS) score as a predictor of 28-day mortality in ED patients with sepsis in The Netherlands, and to compare its performance to C reactive protein (CRP) and lactate. METHODS: This was a historical cohort study in a secondary and tertiary care university hospital. Patients were included if they were seen by an internist in the ED, fulfilled the clinical criteria for sepsis and were admitted to the hospital. Primary outcome was all-cause in-hospital mortality within 28 days. RESULTS: In the 6-month study period, 331 patients were included, of whom 38 (11.5%) died. Mortality varied significantly per MEDS category: ≤4 points (very low risk: 3.1%), 5–7 points (low risk: 5.3%), 8–12 points (moderate risk 17.3%), 13–15 points (high risk: 40.0%), >15 points (very high risk: 77.8%). Receiver operating characteristic (ROC) analysis showed that the MEDS score predicted 28-day mortality better than CRP (area under the curve (AUC) values of 0.81 (95% CI 0.73 to 0.88) and 0.68 (95% CI 0.58 to 0.78), respectively). Lactate was not measured in enough patients (47) for a valid evaluation, but seemed to predict mortality at least fairly (AUC 0.75, 95% CI 0.60 to 0.90). CONCLUSIONS: The MEDS score is an adequate tool for predicting mortality in patients with sepsis in a Dutch internistic ED population. CRP is less useful in this context. Lactate appears to be at least a fair predictor of mortality, but needs to be investigated more systematically in a larger population.

The aim of this study was to investigate farm-level economic and technical factors that are associated with the use of antibiotics on pig farms. Identification of such factors, like farm size and net farm result, may help to increase epidemiological knowledge and to specify farm advice and policy making to reduce inappropriate use of antibiotics. The study used over 300 farm-year records collected during 2004 to 2007 from pig farms in the Netherlands. Data included economic and technical factors as well as antibiotic administration. Data were statistically analyzed for factors associated with antibiotic use, both for fattening pig and sow farms (piglets only), separately. The response variable was the average number of daily dosages per average pig year. Statistical analysis was performed on 16 and 19 potential explanatory factors for the fattening pig and sow farms, respectively. The results showed that, both on the fattening pig and sow farms, the average use of antibiotics increased from 2004 to 2006, but decreased during 2007, but the effect of year was not significant (P> 0.05). Use of antibiotics varied between individual farms. Large farm repeatability for the use of antibiotics in the different years was found. Factors associated (P< 0.05) with the use of antibiotics included: farm system, number of pigs, and population density in the region of the farm (for sow farms only). As these factors are easy to collect and to register, they can be used to specify farm advice and investigation, as well as for policy making. The majority of the technical and economic factors were not significantly (P> 0.05) related to the on-farm use of antibiotics. Therefore, it is recommended to focus future research on the potential role of socioeconomic factors associated with antibiotic use on pig farms.

Harmonization of land use class sets should consider both space and time, as the objective should include harmonization of land use change to analyse environmental processes and problems. Analysis of major class sets reveals that a parametric approach with two parameters may suffice: 'function' that describes land use in an economic context and 'activity' that is defined as the combination of actions resulting in a certain type of product.An example of land use harmonization in Albania illustrates how creation of a reference system based upon the principles of classification, using the synergy between classification and information technology concepts and based upon the 'function' and 'activity' parameters can facilitate harmonization between land use class sets in parallel to achieving land use change harmonization.As not only data quality is of paramount importance, further research is necessary to define quantitative measures to determine harmonization results both at class level and between class sets.

ObjectiveTo investigate the generalizability of the results of a randomized controlled trial on the effectiveness of longterm, highintensity exercises in the rheumatoid arthritis patients in training RAPIT trial by comparing the characteristics of the participants with the nonparticipants.MethodsParticipants and nonparticipants were mailed questionnaires on sociodemographic characteristics, health status, reasons not to participate, and attitudes toward intensive exercise.ResultsThe questionnaires from 892 75 nonparticipants and 299 97 participants were collected. The nonparticipants were slightly older, more often male, and had longer disease duration than the participants. The nonparticipants perceived their disease as more serious, used fewer diseasemodifying antirheumatic drugs, had a lower level of education, and a more negative attitude toward intensive exercise.ConclusionThe results of the RAPIT trial might not be generalizable to the entire target population. To promote participation in longterm, highintensity exercises, health professionals should more actively discuss the potential benefits of exercise with their RA patients while taking into consideration specific factors related to participation.

To investigate the generalizability of the results of a randomized controlled trial on the effectiveness of long‐term, high‐intensity exercises in the rheumatoid arthritis patients in training (RAPIT) trial by comparing the characteristics of the participants with the nonparticipants.Participants and nonparticipants were mailed questionnaires on sociodemographic characteristics, health status, reasons not to participate, and attitudes toward intensive exercise.The questionnaires from 892 (75%) nonparticipants and 299 (97%) participants were collected. The nonparticipants were slightly older, more often male, and had longer disease duration than the participants. The nonparticipants perceived their disease as more serious, used fewer disease‐modifying antirheumatic drugs, had a lower level of education, and a more negative attitude toward intensive exercise.The results of the RAPIT trial might not be generalizable to the entire target population. To promote participation in long‐term, high‐intensity exercises, health professionals should more actively discuss the potential benefits of exercise with their RA patients while taking into consideration specific factors related to participation.

We have investigated frameshift mutations in exonic repeats in the ATR, BRCA1, BRCA2, PTCH, CTCF, Cx26, NuMa and TGFbetaRII genes, using human tumor samples from stomach, esophagus, breast and skin and melanoma, as well as colon cancer and endometrial cancer cell lines (125 samples in total). We developed a sensitive method to detect mutations in the repeats, using the introduction of an artificial restriction site into a repeat. The method detects a single mutant among 10(3) normal genes. Thus, an alteration in a repeated sequence can be detected unambiguously. The (A)(8) repeat of BRCA2 was found mutated in only two of five colon cell lines with microsatellite instability (MI(+)). The ATR gene has an (A)(10) repeat which was altered in two of three MI(+) stomach cancer samples and one of three MI(+) endometrial cell lines. The TGFbetaRII gene [with an (A)(10) repeat] had the maximal frequency of mutations: 10 out of 13 MI(+) samples. At least one sample from all types of cancers, except melanomas, was positive for TGFbetaRII gene mutations. No mutations were found in repeats in the BRCA1, PTCH, CTCF, NuMA and Cx26 genes in any types of tumors examined. In conclusion, our study indicates that repeats were altered only in MI(+) cells and that the mutation frequencies in the genes studied differ among tumor types. Based on these results, we discuss meaningful and meaningless alterations in exonic repeats.

Two BCG preparations produced according to different principles for use as an adjunct to surgery in a comparative clinical trial in melanoma patients have been examined for immunostimulating, toxic, and antitumor activity in various murine and guinea pig models and for immunostimulation in vitro of human lymphocytes. One BCG preparation was prepared in the classic way, i.e., grown as a surface culture and subsequently homogenized in a ball mill, while the other preparation was grown as a dispersed culture.

Transgenic mice carrying heterologous genes directed by a 670-bp segment of the regulatory sequence from the human transferrin (TF) gene demonstrated high expression in brain. Mice carrying the chimeric 0.67kbTF-CAT gene expressed TF-CAT in neurons and glial cells of the nucleus basalis, the cerebrum, corpus callosum, cerebellum, and hippocampus. In brains from two independent TF-CAT transgenic founder lines, copy number of TF-CAT mRNA exceeded the number of mRNA transcripts encoding either mouse endogenous transferrin or mouse endogenous amyloid precursor protein. In two transgenic founder lines, the chloramphenicol acetyltransferase (CAT) protein synthesized from the TF-CAT mRNA was estimated to be 0.10-0.15% of the total soluble proteins of the brain. High expression observed in brain indicates that the 0.67kbTF promoter is a promising director of brain expression of heterologous genes. Therefore, the promoter has been used to express the three common human apolipoprotein E (apoE) alleles in transgenic mouse brains. The apoE alleles have been implicated in the expression of Alzheimer disease, and the human apoE isoforms are reported to interact with different affinities to the brain beta-amyloid and tau protein in vitro. Results of this study demonstrate high expression and production of human apoE proteins in transgenic mouse brains. The model may be used to characterize the interaction of human apoE isoforms with other brain proteins and provide information helpful in designing therapeutic strategies for Alzheimer disease.

The different forms of hyperoxaluria have been discussed, especially the primary hyperoxalurias, autosomal recessive inborn errors of metabolism (type I and II).

Four independent lines of transgenic mice were produced carrying integrated copies of a chimeric gene composed of 3.3 kb of the human haptoglobin 5' regulatory region fused to the CAT (chloramphenicol acetyl transferase) reporter gene. Although the endogenous mouse haptoglobin (Hp) and human haptoglobin (HP) genes express mainly in liver and lung, expression of the human 3.3-kb HP-CAT transgene was not detected until after induction of inflammation and then only in lungs. The results indicated that the transgene maintained the regulatory DNA elements required for lung specific responsiveness to inflammation in vivo but lacked the DNA sequence required for robust expression in liver. The DNA sequence(s) responsible for the normally high level of HP expression in liver either reside outside the 3.3-kb regulatory region of the HP chimeric gene or this region contains a suppressor sequence affecting tissue specific expression in the liver.Copyright 1995, 1999 Academic Press, Inc.

One hundred fifty-seven patients with 208 idiopathic clubfeet were operated on by a single surgeon (K.K.) from 1976 through 1987. Retrospective analysis was performed with a mean follow-up of 7.3 years on 126 patients with 168 operated-on feet. Twelve (7.1%) of 168 feet developed dorsal subluxation of the tarsal navicular. Six (6.3%) of 95 patients developed subluxation with the posterior medial incision. Six (8.2%) of 78 patients developed subluxation with the Cincinnati incision. Seven (58%) of these 12 feet required additional surgery. An anatomic model was constructed to demonstrate how rotation of the navicular alone mimics the radiographic appearance of dorsal subluxation of the navicular. The navicular dorsal subluxation is a common complication and is associated with a high rate of additional corrective surgery. We suggest that dorsal subluxation of the navicular should be redefined as a rotatory deformity with the medial border of the navicular rotating superiorly in the coronal plane. Attention to the position and rotation of navicular at the time of internal fixation may prevent this complication.

Diffuse reflectance laser flash photolysis was successfully employed to study the triplet states of dyes adsorbed on or chemically attached to cotton. This technique, together with the detection of singlet oxygen from dyes on cotton, made it possible to investigate the diffusion of oxygen in cotton fibres and the mobility of dyes adsorbed onto cotton. Absorption by the triplet states of aluminium phthalocyanine chloride and eosin and the phosphorescence of singlet oxygen produced by sensitisation with rose bengal was used to monitor oxygen diffusion and dye mobility by measuring differences in triplet lifetimes, singlet oxygen lifetimes and in signal intensities. It was found that swollen cotton allows diffusion of oxygen in the fibres. However, a noticeable effect on the triplet state of adsorbed aluminium phthalocyanine chloride is only observed if oxygen is removed or added by thorough evacuation or oxygenation of the samples over several days. Singlet oxygen was quenched dynamically by adsorbed dyes, which were found to be immobilised on the fibres, demonstrating that oxygen is mobile within the fibres at the molecular level.

The temperature dependence of the helical pitch in the ferroelectric liquid crystal DOBAMBC near the C*-A transition (Tc) has been measured with a high temperature resolution. The pitch strongly decreases with temperature near the transition but reaches a finite value at Tc. The consistency of the data with different theoretical models is studied.

The yeast Rad4 and Rad23 proteins form a complex that is involved in nucleotide excision repair (NER). Their function in this process is not known yet, but genetic data suggest that they act in an early step in NER. We have purified an epitope-tagged Rad4.Rad23 (tRad4. Rad23) complex from yeast cells, using a clone overproducing Rad4 with a hemagglutinin-tag at its C terminus. tRad4.Rad23 complex purified by both conventional and immuno-affinity chromatography complements the in vitro repair defect of rad4 and rad23 mutant extracts, demonstrating that these proteins are functional in NER. Using electrophoretic mobility shift assays, we show preferential binding of the tRad4.Rad23 complex to damaged DNA in vitro. UV-irradiated, as well as N-acetoxy-2-(acetylamino)fluorene-treated DNA, is efficiently bound by the protein complex. These data suggest that Rad4.Rad23 interacts with DNA damage during NER and may play a role in recognition of the damage.

In Alzheimer′s disease a small fragment of the amyloid protein precursor (APP), called β4, is a characteristic component of senile plaques in brains of affected patients. Efforts to intervene in Alzheimer′s disease include approaches by which APP levels can be decreased in brain. The study described here demonstrates the expression of APP gene in four cell lines that originated from human brain glioblastomas. In one line, HTB 17, APP mRNA level was approximately 25% the APP mRNA found in human brain and 150% that found in human liver. To ascertain whether or not APP expression in HTB 17 cells could be modulated by a cytokine associated with the inflammatory response, cells were cultured in the presence of IL-1β. A significant decrease in APP mRNA accompanied treatment of glioma cells with 1L-1β.

Till now nearly all papers about the Bhabba equation treat equations with a mass term m1. This is merely a matter of mathematical simplicity, because we will see in this work that important physical fields can only be described by a mass term ≠ m 1. The second aim of this publication is to derive a constructive procedure for the calculation of the representations of the Bhabha equation and the tensor and spinor field equations, which are equivalent to them

In this study Agrobacterium tumefaciens transferred DNA (T-DNA) was targeted to a chromosomally introduced lox site in Arabidopsis thaliana by employing the Cre recombinase system. To this end, Arabidopsis target lines were constructed which harboured an active chimeric promoter-lox-cre gene stably integrated in the plant genome. A T-DNA vector with a promoterless lox -neomycin phosphotransferase (nptII) fusion was targeted to this genomic lox site with an efficiency of 1.2-2.3% of the number of random events. Cre-catalyzed site-specific recombination resulted in restoration of nptII expression by translational fusion of the lox-nptII sequence in the integration vector with the transcription and translation initiation sequences present at the target site, allowing selective enrichment on medium containing kanamycin. Simultaneously, the coding sequence of the Cre recombinase was disconnected from these same transcription and translation initiation signals by displacement, aimed at preventing the efficient reversible excision reaction. Of the site-specific recombinants, 89% were the result of precise integration. Furthermore, approximately 50% of these integrants were single copy transformants, based on PCR analysis. Agrobacterium T-DNA, which is transferred to plant cells as a single-stranded linear DNA structure, is in principle incompatible with Cre-mediated integration. Nevertheless, the results presented here clearly demonstrate the feasibility of the Agrobacterium -mediated transformation system, which is generally used for transformation of plants, to obtain site-specific integration.