Restricted immunoglobulin variable region gene usage by hybridoma rheumatoid factors from patients with systemic lupus erythematosus and rheumatoid arthritis.
Rheumatoid factors (RFs) are autoantibodies that are produced by approximately 75% of patients with rheumatoid arthritis (RA). Their role in pathogenesis is not well understood. In this study of 81 human hybridoma IgM antibodies derived from unstimulated peripheral blood B-cells of patients with RA and systemic lupus erythematosus (SLE), we have demonstrated that idiotypes associated with RFs derived from patients with mixed cryoglobulinemia were expressed by approximately 60% of RFs and 6% of IgM antibodies lacking RF activity. The specificity of the RFs for the Fc portion of IgG only (monospecificity) or for Fc and additional self antigens (polyreactivity) was found to correlate with the expression of specific heavy chain associated idiotypes. The VH3 associated RF idiotypes, D12 and B6, were expressed by 0/16 (0%) of monospecific RFs compared with 6/22 (27%) of polyreactive RFs. The predominant use of VH3 was verified by analysis of the expressed Ig with VH family specific anti-peptide antibodies. The light chains expressed by both populations of IgM RFs were found to be predominantly VKIII, both by detection of specific epitopes/idiotypes and V family analysis. This non-random gene usage of both the heavy and light chains suggests that there is a selective expression of V regions in the RF producing B-cells in patients with RA and SLE. We suggest that different antigen-driven, clonal selection events may occur which result in either monospecific RFs or polyreactive RFs.


High Intensity Interval versus Moderate Intensity Continuous Training in Patients with Coronary Artery Disease: A Meta-analysis of Physiological and Clinical Parameters.
Exercise-based cardiac rehabilitation for patients with coronary artery disease (CAD) significantly improves their outcome, although the optimal mode of exercise training remains undetermined. Previous analyses have been constrained by small sample sizes and a limited focus on clinical parameters. Further, results from previous studies have been contradicted by a recently published large RCT. We performed a meta-analysis of published randomised controlled trials to compare high intensity interval training (HIIT) and moderate intensity continuous training (MCT) in their ability to improve patients' aerobic exercise capacity (VO2peak) and various cardiovascular risk factors. We included patients with established coronary artery disease without or without impaired ejection fraction. Ten studies with 472 patients were included for analyses (218 HIIT, 254 MCT). Overall, HIIT was associated with a more pronounced incremental gain in participants' mean VO2peak when compared with MCT (+1.78mL/kg/min, 95% CI: 0.45-3.11). Moderate intensity continuous training however was associated with a more marked decline in patients' mean resting heart rate (-1.8/min, 95% CI: 0.71-2.89) and body weight (-0.48kg, 95% CI: 0.15-0.81). No significant differences were noted in the level of glucose, triglyceride and HDL at the end of exercise program between the two groups. High intensity interval training improves the mean VO2peak in patients with CAD more than MCT, although MCT was associated with a more pronounced numerical decline in patients' resting heart rate and body weight. The underlying mechanisms and clinical relevance of these results are uncertain, and remain a potential focus for future studies.


Inner myometrial laceration: Case report and literature review.
Inner myometrial laceration is a rarely encountered intrapartum uterine trauma, which usually present as post-partum hemorrhage (PPH). The diagnosis can only be confirmed by direct examination of the uterine cavity but it should be suspected in PPH with good uterine contraction. Discussed here are the characteristics and management of the reported cases including two women managed at our center.


Posttranscriptional regulation of ILC2 homeostatic function via tristetraprolin.
Group 2 innate lymphoid cells (ILC2s) are unique in their ability to produce low levels of type 2 cytokines at steady state, and their production capacity is dramatically increased upon stimulation with IL-33. However, it is unknown how constitutive cytokine production is regulated in the steady state. Here, we found that tristetraprolin (TTP/Zfp36), an RNA-binding protein that induces mRNA degradation, was highly expressed in naive ILC2s and was downregulated following IL-33 stimulation. In ILC2s from Zfp36-/- mice, constitutive IL-5 production was elevated owing to the stabilization of its mRNA and resulted in an increased number of eosinophils in the intestine. Luciferase assay demonstrated that TTP directly regulates Il5 mRNA stability, and overexpression of TTP markedly suppressed IL-5 production by ILC2s, even under IL-33 stimulation. Collectively, TTP-mediated posttranscriptional regulation acts as a deterrent of excessive cytokine production in steady-state ILC2s to maintain body homeostasis, and downregulation of TTP may contribute to massive cytokine production under IL-33 stimulation.


Spot diagnosis of aluminium phosphide ingestion: an application of a simple test.
Silver nitrate impregnated paper test was performed with the gastric fluid and in breath in 50 patients of aluminium phosphide poisoning. The test was 100% positive with gastric fluid but was positive in 50% patients in breath. The test is simple, reliable and sensitive method to detect phosphine (PH3) for bed side diagnosis of aluminium phosphide (AIP) ingestion.


Viral adaption of staphylococcal phage: A genome-based analysis of the selective preference based on codon usage Bias.
Given the high therapeutic value of the staphylococcal phage, the genome co-evolution of the phage and the host has gained great attention. Though the genome-wide AT richness in staphylococcal phages has been well-studied with nucleotide usage bias, here we proved that host factor, lifestyle and taxonomy are also important factors in understanding the phage nucleotide usages bias using information entropy formula. Such correlation is especially prominent when it comes to the synonymous codon usages of staphylococcal phages, despite the overall scattered codon usage pattern represented by principal component analysis. This strong relationship is explained by nucleotide skew which testified that the usage biases of nucleotide at different codon positions are acting on synonymous codons. Therefore, our study reveals a hidden relationship of genome evolution with host limitation and phagic phenotype, providing new insight into phage genome evolution at genetic level.


Results, problems and perspectives with the autoregulating pacemaker.
Experimental in vitro testing coupled with animal and clinical studies have shown that a pH sensor may sense acute changes in pH occurring in response to exercise. This signal may in turn be used as a determinant for optimal pacing rate. Further investigation into membrane coating of electrodes and algorithm testing is necessary to maximize sensor response and control.


The Trapezium is Not Necessary: Logical Implications in Treating Basal Joint Arthritis and Pollicization.
It has long been thought that the surgical treatment of osteoarthritis of the first carpometacarpal joint must replicate the normal anatomy. Common sense argues that biomechanical stability can be achieved by a simple ball-and-socket joint obviating complicated ligament reconstructions and trapezium replacements. Our argument is presented and the conclusions are based on the results of a very large series over a long period. A simple trapezium excision arthroplasty of the base of the thumb without ligamentous reconstructions is all that needs to be done to surgically solve painful osteoarthritis of the first carpometacarpal joint. Anything more is overoperating.


Congenital anosmia and emotion recognition: A case-control study.
Patients with anosmia are not able to detect volatile chemicals signaling the presence of infectious and non-infectious environmental hazards, which typically elicit disgust and fear, respectively. Social animals may compensate a loss of olfaction by taking advantage of signals of threat that are produced by their conspecifics. Among humans and other primates, body postures and facial expressions are powerful cues conveying emotional information, including fear and disgust. The aim of the present study was to examine whether humans with agenesis of the olfactory bulb, a rare disorder characterized by congenital anosmia, would be more accurate in recognizing facial expressions of fear and disgust. A total of 90 participants with no history of mental disorder or traumatic brain injury were recruited, including 17 patients with congenital anosmia (10 men, mean age ± standard deviation: 36.5 ± 14.8 years), 34 patients with acquired anosmia (18 men, mean age ± standard deviation: 57.2 ± 11.8 years) and 39 healthy subjects (22 men, mean age ± standard deviation: 36.7 ± 13.2 years). For each patient with congenital anosmia, the agenesis of the olfactory bulb was ascertained through magnetic resonance imaging. Emotion recognition abilities were examined with a dynamic paradigm in which a morphing technique allowed displaying emotional facial expressions increasing in intensity over time. Adjusting for age, education, depression and anxiety, patients with congenital anosmia required similar levels of intensity to correctly recognize fear and disgust than healthy subjects while they displayed decreased error rates for both fear (mean difference [95% confidence interval] = -28.3% [-46.3%, -10.2%], P = 0.003) and disgust (mean difference [95% confidence interval] = -15.8% [-31.5%, -0.2%], P = 0.048). Furthermore, among patients with acquired anosmia, there was a negative correlation between duration of anosmia and the rate of errors for fearful (Spearman's ρ = -0.531, P= 0.001) or disgust (Spearman's ρ = -0.719, P < 0.001) faces recognition. No significant difference was observed for the other primary emotions. Overall, these results suggest that patients with congenital anosmia and long-lasting acquired anosmia may compensate their inability to detect environmental hazards through olfaction by an increased ability to detect fear or disgust as facially expressed by others.


Challenging despair: teaching about women's resistance to violence.
In this article, the author describes an approach to teaching about violence against women that balances discussion of violence with information about women's individual and collective resistance. This strategy addresses two concerns about traditional approaches to this topic: that focusing only on victimization disempowers students and that it provides only a partial view of the reality of violence in women's lives. To address these problems, the author integrates discussion of resistance into the class's working definition of violence, assigned readings, guest speakers, and course assignments. The author concludes with a discussion of the positive effects of this approach.


The potential role of the lymphocyte in fetal wound healing.
Wound healing is a complex process involving a number of different cell populations. Since fetal wounds contain only a small number of mononuclear cells and no fibrosis immunohistochemical techniques using monoclonal antibodies specific to T and B lymphocytes, as well as macrophages, were used to identify the mononuclear cells in fetal wounds and to compare the lymphocyte response with the adult wound healing response. Polyvinyl alcohol sponges were placed subcutaneously in fetal rabbits on day 24 of gestation (then 31 days). Adult rabbits underwent similar implantation. Implants (8 to 14 per group) were harvested 5 days later and examined by H&E and trichrome staining as well as immunohistochemical staining with monoclonal antibody (Mab) specific for rabbit T and B lymphocytes. The sponges in one group of fetal rabbits were impregnated with the polypeptide growth factor, transforming growth factor-beta (TGF-beta), a known regulator of adult tissue repair. Adult wounds showed an intense inflammatory response with extensive collagen deposition; 80% of the infiltrating cellular elements were T lymphocytes. Fetal wounds were less cellular with minimal collagen deposition; 40% of the cells were T lymphocytes. The fetal wounds treated with TGF-beta were intensely cellular and fibrotic with 71% of the cellular infiltrate comprised of T lymphocytes. The increased cellularity of the TGF-beta wounds appears to be primarily the result of increased numbers of T lymphocytes. These findings demonstrate that the fetus mounts an attenuated T lymphocyte response compared to the adult. The increased response of T lymphocytes with TGF-beta suggests that the specific growth factor milieu may account for the fetal response to wounding.(ABSTRACT TRUNCATED AT 250 WORDS)


Excessive discounting of delayed reinforcers as a trans-disease process contributing to addiction and other disease-related vulnerabilities: emerging evidence.
Delay discounting describes the devaluation of a reinforcer as a function of the delay until its receipt. Although all people discount delayed reinforcers, one consistent finding is that substance-dependent individuals tend to discount delayed reinforcers more rapidly than do healthy controls. Moreover, these higher-than-normal discounting rates have been observed in individuals with other behavioral maladies such as pathological gambling, poor health behavior, and overeating. This suggests that high rates of delay discounting may be a trans-disease process (i.e., a process that occurs across a range of disorders, making findings from one disorder relevant to other disorders). In this paper, we argue that delay discounting is a trans-disease process, undergirded by an imbalance between two competing neurobehavioral decision systems. Implications for our understanding of, and treatment for, this trans-disease process are discussed.


The use of expanding ports in laparo-endoscopic single-site surgery may cause more pain: a meta-analysis of randomized clinical trials.
Previous meta-analyses on the clinical outcome after laparo-endoscopic single-site surgery (LESS) versus conventional laparoscopic surgery (CLS) have not revealed any major differences in postoperative pain between the two procedures. This meta-analysis aims to evaluate the difference in postoperative pain between the two procedures, focusing on whether LESS was conducted with a non-expanding port (LESSnonex) or a port expanding (LESSex) within the incision. EMBASE, Medline, PubMed, Science Citation Index Expanded, and Cochrane Central Register of Controlled Trials were searched for randomized clinical trials (RCTs) on LESS versus CLS for general abdominal procedures. Weighted mean difference (WMD) and Odds ratios (OR) were calculated with 95% confidence intervals (CI). A total of 29 RCTs with 2999 procedures were included. Pain (VAS 0-10) 6 h after surgery was significantly lower in the group where LESS was conducted with LESSnonex compared to CLS, WMD=-0.72 (- 1.10 to - 0.33). Pain 18-24 h was significantly higher in the group where LESS was conducted with LESSex compared to CLS, WMD = 0.38 (0.01-0.75). Wound-related complications were significantly more frequent in LESSex procedures compared to CLS, OR = 1.94 (1.03-3.63). The present meta-analysis indirectly indicates that the type of access device that is used for an abdominal LESS procedure may contribute to the development of early postoperative pain as the use of a non-expanding model was associated with a more advantageous outcome. Direct randomized comparison of LESSnonex and LESSex is warranted to confirm if the use of expanding access devices generates more pain and wound complications.


[Neurophysiology of pain in tentorial irritation: description of a case secondary to medulloblastoma].
We report the case of a 32-year-old woman who complained of dull and compressing occipital pain, with unilateral radiation to the left frontal and supraorbital areas (of the sickening type). The radiological, clinical, neurosurgical and neuropathologic investigation disclosed a medulloblastoma bulging and tickening the tentorium cerebelli. This case shows how mechanical stimuli of the structures innervated by the tentorial nerve can cause pain with characteristics of trigeminal and cervical involvement.


Inhibition of fibroblast growth factors.
The potential roles of members of the fibroblast growth factor family in tumor angiogenesis and metastasis and their mechanisms of release from cells are discussed. Furthermore, we review methods of therapeutic targeting of these polypeptides. In particular, we focus on the possibility to inhibit fibroblast growth factors with drugs that mimic heparin-like cellular binding sites and thus can interfere with growth factor receptor recognition. In addition, we discuss antibodies, antisense oligodeoxynucleotides, and ribozymes as approaches to inhibit production and activity of these growth factors.


Myelopathy by lesions of the craniocervical junction in a patient with forestier disease.
The authors report a case of a patient with diffuse idiopathic skeletal hyperostosis (DISH) associated with basilar impression resulting in tetraparesis. To describe neurologic compromise associated with DISH. Neurologic deficits due to DISH are very rare, and only 1 case of basilar impression associated with DISH has previously been reported in the literature. Diagnosis was confirmed by radiograph and MRI, which demonstrated basilar impression associated with a hyperintense signal in the spinal cord on T2-weighted sequences. Transoral resection of the dens associated with posterior occipitocervical fixation was performed during the same anesthesia. Postoperative outcome demonstrated regression of the pyramidal signs without recovery of unassisted walking. Early MRI should be performed in the evidence of spinal cord suffering in patients with DISH. Transoral approach allowed a good decompression of the spinal cord.


Nonholonomic Virtual Constraints for Control of Powered Prostheses Across Walking Speeds.
This paper presents a method to design a nonholonomic virtual constraint (NHVC) controller that produces multiple distinct stance-phase trajectories for corresponding walking speeds. NHVCs encode velocity-dependent joint trajectories via momenta conjugate to the unactuated degree(s)-of-freedom of the system. We recently introduced a method for designing NHVCs that allow for stable bipedal robotic walking across variable terrain slopes. This work extends the notion of NHVCs for application to variable-cadence powered prostheses. Using the segmental conjugate momentum for the prosthesis, an optimization problem is used to design a single stance-phase NHVC for three distinct walking speed trajectories (slow, normal, and fast). This stance-phase controller is implemented with a holonomic swing phase controller on a powered knee-ankle prosthesis, and experiments are conducted with an able-bodied user walking in steady and non-steady velocity conditions. The control scheme is capable of representing 1) multiple, task-dependent reference trajectories, and 2) walking gait variance due to both temporal and kinematic changes in user motion.


The development of the digestive tract in larval European catfish (Silurus glanis L.).
The European catfish, Silurus glanis L., has become an important aqua cultural fish in Croatia, and it is cultivated extensively in ponds in polyculture with carps. The development of the digestive tract in S. glanis was studied with the aim of improving intensive fish production. Research was carried out on S. glanis larval stadium from 1- to 19-day post-hatching (DPH). The main histological methods used were: haematoxylin and eosin staining, periodic acid Schiff staining (PAS), Alcian blue (AB) and toluidin blue staining (TB). A yolk sac was present during the first 5 days (1-5-DPH). During the initial 3-DPH period, there was no trace PAS and AB activity in the digestive tract. Differentiation of the digestive tract began at 3- to 5-DPH. The oesophagus was positive for AB at 5-DPH, PAS and TB after 7-DPH. Differentiation of enterocytes began at 5-DPG and the intestines were complete at 11-DPH. Development of liver and pancreas was also studied. The analysis of data obtained in this study suggests that after 5-DPH catfish larvae have morphologically completed digestive tracts.


Pediatric and child health nursing: A three-phase research priority setting study in Western Australia.
Priority settings are important to plan and direct future research. The aim of this study was to identify the top ten pediatric and child health nursing research priorities from the perspectives of consumers, community, and healthcare professionals in Western Australia. This study used an adapted James Lind Alliance Priority Setting Partnership design with three phases. 1) A planning workshop to inform a survey. 2) A survey using five open-ended-questions distributed between October 2020 and January 2021 through social media and healthcare professionals' emails. Responders' statements were analyzed with content analysis. 3) A consensus workshop to finalize and rank the themes. The planning workshop participants (n = 25) identified gaps such as community child healthcare and confirmed lack of consumer engagement in previous studies. The survey responses (n = 232) generated 911 statements analyzed into 19 themes. The consensus workshop participants (n = 19) merged and added themes, resulting in 16 final themes. The top three ranked themes were: 'access to service', 'mental health and psychological wellbeing', and 'communication'. The research themes are necessarily broad to capture the wide range of issues raised, reflecting the scope of pediatric and child health nursing. The priorities will inform future research to be directed to areas of priority for stakeholders who have often not had a say in setting the research agenda.


Toxicological analysis in a case of endosulfan suicide.
Stomach, small intestine contents, blood, liver, kidney and urine of a 28-years old man, were analyzed for residues of Endosulfan (6, 7, 8, 9, 10, 10-hexachloro-1, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 4, 3-benzo(e)dioxathiepin 3-oxide). The analysis results showed the presence of high concentrations of the two endosulfan isomers in all samples. Since also alcohol was present in all the tissues analyzed, it was concluded that the victim died of a combined endosulfan-alcohol poisoning. No other drugs were found.


Possible epigenetic mechanisms of tumor progression: induction of high-frequency heritable but phenotypically unstable changes in the tumorigenic and metastatic properties of tumor cell populations by 5-azacytidine treatment.
Treatment of a variety of highly tumorigenic mouse lines in vitro with chemical mutagens, such as ethyl methane sulfonate (EMS) or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), can result in extraordinarily high frequencies (sometimes in excess of 90%) of strongly immunogenic clones unable to grow progressively in normal syngeneic hosts. These clones will, however, grow in immunosuppressed hosts and gradually regain tumorigenic ability in normal mice if maintained in long-term (several months-1 year) culture, i.e., they are often phenotypically unstable. These features--phenotypic drift and high frequency--make it unlikely that point mutations are the underlying mechanism involved in the generation of the variants. Results presented here demonstrate that these observations can be reproduced on the same tumor lines using 5-azacytidine--an analogue of cytidine which can be incorporated into DNA causing subsequent extensive hypomethylation of cytosine residues in the absence of any significant mutagenic effects. Furthermore, 5-azacytidine treatment of a nonmetastatic mouse mammary tumor led to the emergence of a small number of heritable but unstable tumor clones capable of spontaneous metastatic spread. Because it is known that DNA hypomethylation can lead to transcriptional activation of normally silent genes, that altered methylation patterns can be somatically replicated with a high but not perfect fidelity, and that mutagens can cause DNA hypomethylation, we propose that DNA hypomethylation followed by de novo methylation represents a plausible mechanism to account not only for the induction of the nontumorigenic variants but for a number of aspects of tumor progression and tumor heterogeneity, as well. In particular, we refer to heritable phenotypic alterations in tumor cell populations which occur at very high frequency but which are not necessarily stable over very long periods of time.


Ferulago angulata extract ameliorates epididymal sperm toxicity in mice induced by lead and diazinon.
The potential toxicity that results from environmental xenobiotics is not completely known. Increasing levels of heavy metals and the use of organophosphate pesticides (OPs) and their co-existence in the environment could be associated with an increasing incidence of male reproductive system disorders in humans and animals. Ferulago angulata is a dietary source of phenolic compounds with reported health benefits. This study was conducted to investigate whether an extract of Ferulago angulata could protect adult male NMRI mice against reproductive toxicity induced by lead acetate (PbAc), diazinon (DZN), or PbAc + DZN. Adult male NMRI mice were exposed to either 0.5% PbAc in drinking water, DZN (3 mg/kg/day, intraperitoneal [i.p.] injection), or PbAc + DZN in the presence or absence of 400 mg/kg/day Ferulago angulata hydroalcoholic extract (FAE) that was administered via gavage for 6 weeks. Chronic exposure to PbAc, DZN, and PbAc + DZN decreased sperm quality, sperm chromatin maturity and integrity, increased oxidative stress and lipid peroxidation, and could reduce male fertility indices. Co-administration of FAE could reduce these negative effects. The Ferulago angulata extract should be considered as a useful natural extract for the treatment of male infertility, especially in males exposed to conditions which induce reproductive toxicity.


Review of historic article: Ebashi, S & Endo, M. 1968 Calcium Ion and Muscle Contraction. Progress in Biophysics and Molecular Biology, 18, 123-183.
The 1968 review article on Calcium ion and muscle contraction by Setsuro Ebashi and Makoto Endo is one of the highest cited in the journal since it was required reading in the early days of understanding what triggers contraction of the myofilaments. It correctly identified the major steps in excitation-contraction coupling and still inspires mathematical models of muscle activity today. It also successfully identified the role of troponin.


Effects of retinal eccentricity and acuity on global-motion processing.
The present study assessed direction discrimination with moving random-dot cinematograms at retinal eccentricities of 0, 8, 22, and 40 deg. In addition, Landolt-C acuity was assessed at these eccentricities to determine whether changes in motion discrimination performance covaried with acuity in the retinal periphery. The results of the experiment indicated that discrimination thresholds increased with retinal eccentricity and directional variance (noise), independent of acuity. Psychophysical modeling indicated that the results for eccentricity and noise could be explained by an increase in channel bandwidth and an increase in internal multiplicative noise.


Functional differences between peroxidase compound I and the cytochrome P-450 reactive oxygen intermediate.
A series of seven hemeproteins, cytochromes P-450LM2, P-450LM4, and P-420LM2, horseradish peroxidase, chloroperoxidase, catalase, and metmyoglobin, as well as hemin were tested for their ability to catalyze a set of five oxidative reactions. These reactions were a typical peroxidative reaction (oxidation of pyrogallol to purpurogallin) and three characteristic P-450 reactions (aliphatic hydroxylation, aromatic hydroxylation, and olefinic epoxidation). In addition, the ability to decarboxylate a peroxyacid was measured. All hemeproteins were able to carry out peroxidation, but three (horseradish peroxidase, chloroperoxidase, and catalase) were much better catalysts than the others. Only the P-450 enzymes were competent catalysts for the hydroxylation and epoxidation reactions. Furthermore, the decarboxylation reaction was strictly limited to the P-450 enzymes, establishing it as a new, unique P-450 activity. Since the decarboxylation of peroxyacids is diagnostic of peroxide homolysis, these results indicate a fundamentally different manner of processing of peroxides by cytochrome P-450 than by the peroxidases. Thus, the possibility of close similarity of reactive oxygen intermediates in the two series is called into question.


TIMP-3 deficiency in the host, but not in the tumor, enhances tumor growth and angiogenesis.
Tumor cells, stromal cell compartment and the extracellular matrix (ECM) together generate a multifaceted tumor microenvironment. Matrix metalloproteinases and their tissue inhibitors (TIMPs) provide a means for tumor-stromal interaction during tumorigenesis. Among TIMPs, TIMP-3 is uniquely localized to the ECM and is frequently silenced in human cancers. Here, we asked whether the absence of TIMP-3 in the tumor cell or the host affects the process of tumorigenesis. Timp-3(-/-) ES-cell clones were generated and used to develop teratomas in nude mice. Timp-3(-/-) teratomas showed similar tumor take, growth, and angiogenesis compared to timp-3(+/+) teratomas. To study the effect of TIMP-3 ablation in the host stroma, we measured the growth kinetics of subcutaneous B16F10 melanomas in timp-3(-/-) and wild-type littermates. Tumors grew significantly faster in timp-3(-/-) than in wild-type mice and their CD31 content was significantly higher indicating increased angiogenesis. Augmented angiogenesis in timp-3(-/-) mice was directly tested using Matrigel plug and Gelfoam assays. In response to FGF-2, timp-3(-/-) endothelial cells invaded more efficiently, leading to enhanced formation of functional blood vessels. Thus, TIMP-3 deficiency in the host, but not in the tumor per se, leads to enhanced tumor growth and angiogenesis. TIMP-3 located within the tumor microenvironment inhibits tumorigenesis.


New proposed cut-off of waist circumference for central obesity as risk factor for diabetes mellitus: Evidence from the Indonesian Basic National Health Survey.
Waist circumference (WC) measurement is practical to define central obesity. However, WC cut-off point might be differ based on different race or ethnicity. This study aims to analyze the optimal WC cut-off point to identify T2DM risk among Indonesian population. We analyzed the secondary data of national based cross-sectional study of the Indonesian Basic National Health Research 2013, comprising 24,660 adults aged ≥ 18 years who were assessed for fasting plasma glucose (FPG) and oral glucose tolerance test (OGTT). The new proposed cut-off point for WC was calculated using ROC curve analysis and Youden index. The odds ratio of having T2DM was calculated using logistic regression analysis. Increased WC was associated with worsening dysglycemia status among men and women (p<0.001). The optimal cut-off point of WC for detecting T2DM from ROC analysis was 76 for men and 80 for women. Based on this WC cut-off point, the odds ratio for having T2DM was 1.64 [95% CI 1.45-1.86, p<0.01] for men and 1.90 [95% CI 1.71-2.11 p<0.01] for women. The newly proposed WC cut-off point of 76 for men and 80 women can be used to screen the risk of T2DM among Indonesian population.


Pituitary-adrenal axis and peripheral cortisol metabolism in obese patients.
A close relationship between adiposity and increased cortisol levels is well established in patients with endogenous hypercortisolism. Nevertheless, hypothalamic-pituitary-adrenal (HPA) axis regulation in overweight subjects is still a matter of concern. We studied free cortisol (urinary free cortisol, UFC and late night salivary cortisol, LNSC), pituitary feedback (serum cortisol after 1 mg dexamethasone suppression test, 1 mg DST) and peripheral cortisol metabolism (urinary cortisol to cortisone ratio, F/Eratio) in a large series of overweight subjects without Cushing's Syndrome. We considered 234 patients divided in 5 BMI classes, matched for age and gender (BMI ≤ 25 kg/m2n = 38; 25-30 n = 58; 30-35 n = 52; 35-40 n = 52; >40 n = 34). UFC, LNSC and urinary F/Eratio were assessed with LC-MS. We collected 183 LNSC, 176 UFC, 152 1 mg DST and 64 F/Eratio tests. UFC levels were higher in lean subjects, and they decreased according to the BMI classes (p = 0.022). Non-suppressed cortisol levels (>50 nmol/L) after 1 mg DST were observed especially in patients with normal weight or mild obesity. Patients with BMI ≥ 35 kg/m2 revealed a reduced F/Eratio (0.39 vs. 0.61, p = 0.006). The specificity of tests (false positive results) was higher considering 1 mg DST or UFC in obese patients, on the contrary impaired cortisol rhythm (LNSC above normality) was observed in 47 subjects, irrespective of weight. Overweight and obese subjects are characterised by an original regulation of HPA axis (reduced UFC levels, increased suppression after 1 mg DST) and peripheral cortisol metabolism (reduced F/Eratio), suggesting an effort to counteract hypercortisolism.


Calcium uptake by sarcoplasmic reticulum and its modulation by cAMP-dependent phosphorylation in normal and failing human myocardium.
ATP-dependent, oxalate-supported Ca2+ uptake by cardiac sarcoplasmic reticulum was examined in microsomes prepared from left-ventricular free wall myocardium obtained from the explanted failing hearts of transplant recipients with idiopathic dilated cardiomyopathy and the non-failing hearts of kidney donors for whose hearts no suitable recipients were available. There were no significant differences between the two groups with respect to values for Vmax, K0.5 (for Ca2+) or nHill of basal Ca2+ uptake. The stimulation of Ca2+ uptake associated with cAMP-dependent phosphorylation of phospholamban could be reproduced by incubation of microsomes with a monoclonal antibody to phospholamban. Stimulation resulted from a decrease in K0.5, with no changes in Vmax or nHill. The magnitude of stimulation of Ca2+ uptake following incubation with anti-phospholamban monoclonal antibody was identical in preparations from normal and failing hearts. Finally, sarcoplasmic reticulum-associated cGMP-inhibited cAMP phosphodiesterase activity in these preparations was characterised. Measurement of steady-state kinetics and pharmacologic sensitivity indicated that this activity was functionally homogeneous. Preparations from failing and non-failing hearts did not differ with respect to either values for Vmax and Km or susceptibility to inhibition by the cilostamide derivative OPC 3911. These observations indicate that abnormalities in the regulation of intracellular [Ca2+] in failing human myocardium cannot be ascribed to changes in the level or function of the Ca(2+)-transporting ATPase, phospholamban or cGMP-inhibited cAMP phosphodiesterase in the sarcoplasmic reticulum.


The mutational landscape of the SCAN-B real-world primary breast cancer transcriptome.
Breast cancer is a disease of genomic alterations, of which the panorama of somatic mutations and how these relate to subtypes and therapy response is incompletely understood. Within SCAN-B (ClinicalTrials.gov: NCT02306096), a prospective study elucidating the transcriptomic profiles for thousands of breast cancers, we developed a RNA-seq pipeline for detection of SNVs/indels and profiled a real-world cohort of 3,217 breast tumors. We describe the mutational landscape of primary breast cancer viewed through the transcriptome of a large population-based cohort and relate it to patient survival. We demonstrate that RNA-seq can be used to call mutations in genes such as PIK3CA, TP53, and ERBB2, as well as the status of molecular pathways and mutational burden, and identify potentially druggable mutations in 86.8% of tumors. To make this rich dataset available for the research community, we developed an open source web application, the SCAN-B MutationExplorer (http://oncogenomics.bmc.lu.se/MutationExplorer). These results add another dimension to the use of RNA-seq as a clinical tool, where both gene expression- and mutation-based biomarkers can be interrogated in real-time within 1 week of tumor sampling.


A comparatively optimization of dosages of oxidation agents based on volatile solids and dry solids content in dewatering of sewage sludge.
Organic content of sludge is a major factor influencing its dewaterability. Conditioning sewage sludge with oxidation reagents (Fenton's reagent or Fe2+/persulfate) can effectively improve sludge dewaterability. In traditional conditioning process of sewage sludge, the optimization of conditioner dosage were commonly based on volume of sludge (referred as mg/L) or mass of dry solid (DS) of sludge (referred as mg/g DS). However, inconsistency of the oxidation dose mode existed for different sewage sludge sources. In this study, sludge samples of different volatile solids (VS) contents were used to derive optimal dosages of Fenton's reagent and Fe2+/persulfate, using the response surface methodology (RSM). For the case of Fenton's reagent, the optimal dosages of Fe2+ and H2O2 were 107-110 mg/g VS and 86-88 mg/g VS, respectively. For Fe2+/persulfate, the optimal dosages of Fe2+ and persulfate were 49 mg/g VS and 269-271 mg/g VS, respectively. The optimal dosages of the oxidation reagents based on VS contents were proved to be consistent and effective for different sewage sludge with different organic matter contents from different wastewater treatment plants (WWTPs). In contrast, the optimal dosages of oxidation reagents, based on DS, fluctuated significantly for different sludge sources. Furthermore, sewage sludge dewaterability was significantly related to the degradation and the content of loosely bound extracellular polymeric substances (LB-EPS) in the organic matters of conditioned sludge (R2 > 0.9, p < 0.01). Thus, the improvement of sludge dewaterability could be related with the destruction of the VS in sludge with the conditioners of oxidant reagent. It indicated that optimization of oxidation reagent based on VS content is more plausible than that of based on DS content for different sewage sludge with different organic matter contents.


Seroepidemiology of Toxoplasma gondii infection in pregnant women in a public hospital in northern Mexico.
Toxoplasma gondii (T. gondii) infection in pregnant women represents a risk for congenital disease. There is scarce information about the epidemiology of T. gondii infection in pregnant women in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic, clinical and behavioural characteristics in a population of pregnant women of Durango City, Mexico. Three hundred and forty three women seeking prenatal care in a public hospital of Durango City in Mexico were examined for T. gondii infection. All women were tested for anti-T. gondii IgM and IgG antibodies by using IMx Toxo IgM and IMx Toxo IgG 2.0 kits (Abbott Laboratories, Abbott Park, IL, USA), respectively. Socio-demographic, clinical and behavioural characteristics from each participant were also obtained. Twenty one out of the 343 (6.1%) women had IgG anti-T. gondii antibodies. None of the 343 women had IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with living in a house with soil floor (adjusted OR = 7.16; 95% CI: 1.39-36.84), residing outside of Durango State (adjusted OR = 4.25; 95% CI: 1.72-10.49), and turkey meat consumption (adjusted OR = 3.85; 95% CI: 1.30-11.44). Other characteristics as cat contact, gardening, and food preferences did not show any association with T. gondii infection. The prevalence of T. gondii infection in pregnant women of Durango City is low as compared with those reported in other regions of Mexico and the majority of other countries. Poor housing conditions as soil floors, residing in other Mexican States, and turkey meat consumption might contribute to acquire T. gondii infection.


Long-term safety of anti-TNF adalimumab in HBc antibody-positive psoriatic arthritis patients: a retrospective case series of 8 patients.
Immunosuppressive drugs commonly used in the treatment of psoriatic arthritis make patients more susceptible to viral, bacterial, and fungal infections because of their mechanism of action. They not only increase the risk of new infections but also act altering the natural course of preexisting infections. While numerous data regarding the reactivation of tuberculosis infection are available in the literature, poor information about the risk of reactivation or exacerbation of hepatitis viruses B and C infections during treatment with biologics has been reported. Furthermore, reported series with biological therapy included short periods of followup, and therefore, they are not adequate to verify the risk of reactivation in the long-term treatment. Our study evaluated patients with a history of hepatitis B and psoriatic arthritis treated with adalimumab and monitored up to six years. During the observation period, treatment was effective and well tolerated in all patients, and liver function tests and viral load levels remained unchanged.


Parasitic ion-implantation produced by a Kaufman-type ion source used for planar etching of surfaces.
Total reflection X-ray fluorescence spectrometry (TXRF) and photoelectron spectroscopy (XPS) were used to characterize ion-etched surfaces of Si wafers, a quartz glass and vacuum-metallized multilayer on a Si wafer. These samples were first treated with low kinetic energy ions of a broad beam Kaufman-type ion source. By repeating this method of sputtering, layers can be removed stepwise and in combination with surface analysis by TXRF a multielement depth profiling can be carried out. The depth resolution is of the order of 3 nm. Unfortunately, ion-etching leads to uncontrollable ion implantation and thus to a contamination of the samples with some 'parasitic' elements which may disturb the multielement characterization of the original surfaces. XPS with Ar sputtering was used to confirm the implantation of interfering elements in the freshly ion-etched samples.


Methods for estimating the incidence of primary infection in pregnancy: a reappraisal of toxoplasmosis and cytomegalovirus data.
Accurate incidence information is required to plan and evaluate screening programmes which have been proposed for the detection of primary toxoplasmosis and cytomegalovirus infection in pregnancy. Appropriate statistical methods are described for deriving incidence rates and their confidence intervals from three types of data: change in age-specific seroprevalence, seroconversion, and IgM studies. These methods are applied to seven published studies on toxoplasmosis and cytomegalovirus carried out in the UK. In these publications only one estimate of the infection rate per pregnancy was correctly derived, and none were accompanied by confidence intervals. Using the proposed methods, most estimates of the primary toxoplasmosis rate in these studies were between 2.5 and 5.5 per 1000 pregnancies, compared to the 2 per 1000 usually cited. Most cytomegalovirus incidence estimates were between 4 and 10 per 1000 pregnancies.


Cell cycle-dependent multiplication of avian adenoviruses in chicken embryo fibroblasts.
Propagation of CELO virus employing confluent monolayers of chicken enbryo fibroblasts (CEF) yielded virus titers one to two logs lower than those from confluent chicken kidney (CK) cells. An enhancement of virus production in CEF as measured by plaque formation was obtainedby infectng cultures in the growing non confluent state. Measurements of 3H-thymidine incorporation revealed a positive correlation between the DNA synthesis of CEF cultures at the time of inoculation and the amount of progeny virus, whereas in the CK-CELO-system no such relation was observed. Requirement of replicative fibroblasts for CELO multiplication was also demonstrated by comparison of virus replication in synchronized stationary and serum stimulated CEF cells. In stationary CEF cells arrested in the G1 phase of the cell replication cycle by serum deprivation and infected withe CELO virs, no cytopathic effect could be observed, and only very low amounts of virus were produced. But 24 hours after release of these cells for growth by serum stimulation a logarithmic rate of virus multiplication and a complete CPE occurred. Infection of synchronized CEF cultures at different stages of the cell cycle revealed that CELO multiplication was correlated with the S phase of the infected cell. In synchronized CELO infected CEF cultures viral DNA synthesis started 12 to 14 hours after growth stimulation when cells were near the end of the S phase. In contrast, no viral DNA synthesis could be measured in growth arrested CELO infected CEF cells, when cellular DNA synthesis was low. Therefore not only production of infectious virus but also viral DNA synthesis is correlated with events during the S phase of the infected CEF cell.


Chemometric study of Andalusian extra virgin olive oils Raman spectra: Qualitative and quantitative information.
Authentication of extra virgin olive oil (EVOO) is an important topic for olive oil industry. The fraudulent practices in this sector are a major problem affecting both producers and consumers. This study analyzes the capability of FT-Raman combined with chemometric treatments of prediction of the fatty acid contents (quantitative information), using gas chromatography as the reference technique, and classification of diverse EVOOs as a function of the harvest year, olive variety, geographical origin and Andalusian PDO (qualitative information). The optimal number of PLS components that summarizes the spectral information was introduced progressively. For the estimation of the fatty acid composition, the lowest error (both in fitting and prediction) corresponded to MUFA, followed by SAFA and PUFA though such errors were close to zero in all cases. As regards the qualitative variables, discriminant analysis allowed a correct classification of 94.3%, 84.0%, 89.0% and 86.6% of samples for harvest year, olive variety, geographical origin and PDO, respectively.


Thermodynamic analysis of calcium binding to frog parvalbumin.
The enthalpy of calcium binding to frog parvalbumin (Rana temporaria isoenzyme IVb, pI 4.75) has been measured by microcalorimetry. The reaction is exothermic; the heat of the reaction in 100 mM KCl, 50mM Tris, pH 8.0 at 12 degrees C is -19 kJ (mol site)-1 and -33 kJ (mol site)-1 in the presence of 1 mM magnesium. The shape of the titration curve indicates that the properties of the two calcium binding sites are different. The thermodynamic parameters measured for frog parvalbumin are compared with those of related parvalbumins from carp and whiting.


Ergot alkaloids: Quantitation and recognition challenges.
Bread, flour, infant formula and baby food samples (n=109, from which n=54 made of or containing rye), collected in 2001, 2003, and 2005, were analysed for ergot alkaloids. Samples were extracted using acidic conditions and the extracts subjected to an automated solid-phase clean up using combined cation exchange/reversed-phase sorbent cartridges (Oasis-MCX). Subsequent chromatographic separation and analysis was performed by liquid chromatography (LC) with fluorescence detection (FLD) and by LC with mass spectrometric detection (MS/MS). The ergot alkaloid (EAs) content of a sample was defined as the sum of the 16 alkaloids ergometrin(in)e, ergosin(in)e, ergotamin(in)e, ergostin(in)e, ergocornin(in)e, α-ergocryptin(in)e, β-ergocryptin(in)e and ergocristin(in)e. Comparability of results obtained by LC-FLD and LC-MS/MS was satisfactory, but varied for different alkaloids. The use of dihydro-ergocristine as an internal standard considerably improved the reliability of analytical data from LC-MS/MS. Compared with earlier data (Baumannet al., 1985) for median levels of ergot alkaloids in rye flour (140 ng/g) and bread (21.3 ng/g) from Switzerland, the median values for ergot alkaloids in rye flour collected in 2001 (n=13) and in 2005 (n=2) were 172 ng/g and 160 ng/g, respectively. The median values for bread (fresh weight) collected in 2001 (n=14), 2003 (n=7), and 2005 (n=2) were 87 ng/g, 120 ng/g, and 156 ng/g, respectively. Low levels of ergot alkaloids were also found in wheat products and in some infant formulae and baby foods containing rye. By additional LC-MS/MS experiments, the possible natural occurrence of ergot congeners containing the 9,10-unsaturated ergoline cation (m/z=223) was investigated. In a few samples, ergovalin(in)e was tentatively identified by these means.


Prognostic Factors in Non-occlusive mesenteric ischemia: A pragmatic pre-operative score for the prediction of 28-day mortality.
The prognosis of critical ill patients with non-occlusive mesenteric ischemia (NOMI) is poor and not fully understood. We aimed to determine preoperative factors associated with 28-day mortality in NOMI. Variables associated with 28-day mortality were entered into a multivariate cox regression model and were used to compute a NOMI mortality score. 154 patients were included. The 28-day mortality rate was 56%. Multivariable analyses including variables at the time of the CT identified three variables (i.e. lactates > 7 mmoL/l, prothrombin rate <60% and kidney infarction), included in a simple score. Among the study population, the probability of 28-day mortality was 26% (11/42), 54% (26/48), 77% (23/30) and 100% (21/21) for a survival score of 0, 1, 2 and 3, respectively. A simple score combining these three variables, calculated preoperatively, was able to accurately predict 28-day mortality and might help to avoid futile laparotomies.


Comprehensive characterization of phytochemicals and biological activities of the Italian ancient apple 'Mela Rosa dei Monti Sibillini'.
This study was carried out to characterize extracts from nine samples of the apple 'Mela Rosa dei Monti Sibillini' (MR) and to assess the antioxidant and anti-inflammatory activities. The extracts were analysed by High Performance Liquid Chromatography coupled with photodiode array detector and mass spectrometry (HPLC-DAD-MS) for 20 phytochemicals. The extracts from the lyophilized material (ELM) were richer in polyphenolic compounds than the dried ones (EDM). The MR extracts contained noteworthy amounts of the investigated analytes compared to one sample of the commercial varieties Annurca, Golden Delicious and Granny Smith used as reference. Principal component analysis (PCA) revealed that the part of the fruit seems to have a significant influence on the chemical composition of the final extract; thus, the peel extracts exhibited higher levels of phenolic compounds, especially epicatechin, procyanidin B2 and phloridzin, and triterpenes than the pulp ones. In general, the lyophilized material showed higher antioxidant activity than the dried material. The strong antioxidant capacity of the MR has also been revealed by the DPPH, ABTS, FRAP and Folin-Ciocalteau assays. The ELM of MR significantly reduced reactive oxygen species in lipopolysaccharide (LPS)-activated mouse brain microglia cells (BV-2 cells). Real-time polymerase chain reaction (RT-PCR) analysis demonstrated that the EDM and ELM of MR were effective in reducing pro-inflammatory cytokines and enzymes in BV-2 and peripheral blood mononuclear cells (PBMC). These results contribute to the exploitation of this ancient variety as a source of nutraceuticals.


Defined, serum-free culture conditions for the GM-micro-clonogenic assay using agar-capillaries.
Culture conditions were determined and optimised for the serum-free growth of granulocyte-macrophage (GM) colonies, derived from mouse bone marrow cells, in agar-containing glass capillaries. The standard medium is DMEM/F-12 (1:1) mixture containing per ml: 10 mg BSA, 35 micrograms transferrin, 20 micrograms soybean lipids and 5 micrograms insulin. In contrast to previous attempts by others, GM-colony yield in serum-free medium was found to be equal to that in serum-containing medium (around 25 colonies/capillary) and to correlate satisfactorily with the cell density at seeding.--The role of polyamine oxidases in cell-proliferation experiments could be studied by this microclonogenic assay without interference from any type of serum.


Mindfulness in Motion and Dietary Approaches to Stop Hypertension (DASH) in Hypertensive African Americans.
Hypertension increases the risk of developing Alzheimer's disease or related dementias. This pilot study's purpose was to examine the feasibility and acceptability of a novel intervention, Mindfulness in Motion (MIM) and Dietary Approaches to Stop Hypertension DASH (MIM DASH), to improve diet, mindfulness, stress, and systolic blood pressure (BP) in older African Americans with mild cognitive impairment (MCI) and hypertension. Cluster randomized controlled trial. Intergenerational community center in a large metropolitan area. African Americans with MCI and hypertension. Participants were divided into six groups randomized 1:1:1 to the MIM DASH group, attention only (non-hypertensive education) group, or true control group. The MIM DASH and attention only interventions were delivered in 8-weekly 2 hour group sessions. MIM included mindful movements from chair/standing, breathing exercises, and guided meditation. The DASH component used a critical thinking approach of problem solving, goal setting, reflection, and self-efficacy. The true control group received a DASH pamphlet at the end. Feasibility was tracked through enrollment and attendance records; acceptability was assessed through interviews. Blood pressure was measured using the Omron HEM-907XL Monitor. Dietary intake was measured by DASH-Q. Mindfulness was measured by the Cognitive and Affective Mindfulness Scale. Stress was measured by the Perceived Stress Scale. MCI was determined using the Self-Administered Gerocognitive Examination. Data were collected at baseline and 3-months. Median session attendance was six for the MIM DASH group and six for the attention only group. There were no changes in diet, mindfulness, or stress. There was a clinically significant reduction in systolic BP in the MIM DASH group (-7.2 mmHg) relative to the attention only group (-.7), and no change between the MIM DASH and true control groups. Results indicate that the MIM DASH intervention was feasible and culturally acceptable in African Americans with hypertension and MCI.


[The organogenic development of the human peroneal muscle of the IVth digit from the peroneal muscle of the Vth digit and the short peroneal muscle from the viewpoint of evolutive myology].
The preliminary author's investigations revealed, that the mm. peronei digitorum group in Mammalia (including Man) underwent its own morphological evolution. The author failed to establish the facts that indicated the homology between the above mentioned muscles and m. extensor digitorum brevis. But, at present due to the lack of detailed determinations of organogenesis of mm. peronei digitorum and of m. peroneus brevis, it was not possible to prove the above mentioned indications. The observations on organogenesis of the respective muscles, carried out at present upon 13 human embryos with 12 to 32 mm crown-rump length and 5 fetuses with 35 to 65 mm crown-rump length, revealed that, mm. peronei digitorum primordium initiated its development as a muscle of two fingers (m. peroneus digiti IV. et m. peroneus digiti V), passed through its one-finger form (m. peroneus digiti V) and usually terminated with its complete reduction.


Potency of L-364,718 as an antagonist of the behavioral effects of peripherally administered cholecystokinin.
A new antagonist of the peripheral cholecystokinin receptor, L-364,718, was found to block the reductions in food intake and exploratory activity induced by intraperitoneal administration of cholecystokinin octapeptide sulfate. L-364,718 significantly reversed the cholecystokinin-induced reduction in feeding at doses of 10 micrograms/kg - 10 mg/kg i.p. L-364,718 significantly reversed the cholecystokinin-induced reduction in exploratory activity at doses of 500 ng/kg - 10 mg/kg i.p. The time course of antagonist activity of L-364,718 was immediate to 90 minutes after intraperitoneal administration. L-364,718 had no significant effect on food intake or exploratory activity when administered alone, over the dose range of 100 ng/kg-10 mg/kg i.p. This compound appears to be at least one hundred times more potent than proglumide or benzotript as an antagonist of the behavioral effects of peripherally administered cholecystokinin.


Magnetic sensor data acquisition for three-dimensional ultrasound of the orbit.
Three-dimensional (3D) B-mode ultrasound imaging is now possible using a magnetic sensor data acquisition system to localise data points, allowing free-hand scanning. We report the first use of this freehand scanning system for insonation of the orbit. We insonated 10 orbits in 5 volunteers (all men; mean age 37 years) with an Acuson 128XP ultrasound system and a 7.5 MHz transducer. Power-Doppler was used to image vascular structures. Data were acquired using a new magnetic sensor system with detectors mounted on the transducer. Free-hand scanning was done using sweeping or fan-like and linear movements over 20-30 s to cover the orbit, with retrospective measurements of orbital structures. A 3D reconstruction was performed via an external workstation. All orbits were adequately imaged by one data acquisition. Selected intraorbital structures were identified and their course followed, including optic nerve and central vessels, as well as short and long ciliary vessels. The mean transverse area of the optic nerve was 5.6 +/- 1.1 mm2; the diameter of the optic nerve was 3.0 +/- 0.3 mm. The lateral rectus muscle was clearly seen, with a mean diameter of 4.9 +/- 0.3 mm, at the level of optic nerve head. Three-dimensional ultrasound of the orbit allows imaging of the location, course and relationships of intraorbital vessels, the optic nerve and the lateral rectus muscles with one data acquisition, as well as quantitative measurements not possible with 2D images alone. This method allows rapid, free-hand data acquisition, with a multitude of potential clinical applications.


Relations between environment, migration and reproduction in a seed bug, Neacoryphus bicrucis (Say) (Heteroptera: Lygaeidae).
The seed feeding bug Neacoryphus bicrucis migrates by flight, often at great altitudes, and evidently is able to disperse over long distances. Non-diapausing females provided with ample food and mates histolyze their flight muscles soon after adult eclosion, and thus undertake very little flight. If food density is below a certain threshold flight ability is retained at a constant high level through most of adult life, while egg production becomes a linear function of the logarithm of food density. Bugs totally starved from adult eclosion onwards survive for a month or more with only a minor decrease in flight ability. Females also retain their flight potential when subjected to lack of mates or diapause-inducing photoperiodic conditions. Most males are able to fly throughout their adult lives under all conditions studied, but the tendency to fly is enhanced by lack of food or mates.


Isolation of multidrug-resistant Stenotrophomonas maltophilia from cultured yellowtail (Seriola quinqueradiata) from a marine fish farm.
Six strains of multidrug-resistant Stenotrophomonas maltophilia were isolated from cultured yellowtail. The strains were divided into two clusters based on the 16S rRNA genes, and all of them contained L1 metallo-beta-lactamase and L2 beta-lactamase genes. Differences in the intercluster divergence between the lactamase genes suggest that horizontal transfer of the genes occurred.


Design and Methodology of the Longitudinal Aging Study in India-Diagnostic Assessment of Dementia (LASI-DAD).
To provide high-quality data on older adults in India that will enable an in-depth study of late-life cognition and dementia in India and cross-country analysis of risk factors for cognitive aging and dementia. The Longitudinal Aging Study in India (LASI) is a nationally representative survey of health, economic, and social well-being of the Indian population aged 45 and older. Its large sample of more than 70,000 older adults represents not only the country as a whole but also each state. LASI-Diagnostic Assessment of Dementia (DAD) is an in-depth study of late-life cognition and dementia, drawing a subsample of over 3,000 LASI respondents aged 60 and older. Participants were interviewed at home or in a participating hospital according to their preferences. Adults aged 60 and older (N = 3,224), along with 3,191 informants. Respondents underwent a battery of cognitive tests, and informants were interviewed about their cognitive and health conditions. A common set of cognitive tests was selected to enable international comparisons, and additional cognitive tests suitable for illiterate and innumerate populations were also selected. Rich data on risk factors of dementia were collected through health examination, venous blood assays, and genotyping. The response rate was 82.9%, varying across sex, education, and urbanicity. Data are available to other researchers. LASI-DAD provides an opportunity to study late-life cognition and dementia and their risk factors in the older population in India and to gain further insights through cross-country analysis by pooling data from its international sister studies. J Am Geriatr Soc 68:S5-S10, 2020.


Convergent tip-to-base widening of water-conducting conduits in the tallest bryophytes.
Tip-to-base conduit widening is considered a key mechanism that enables vascular plants to grow tall by decreasing the hydraulic resistance imposed by increasing height. Widening of hydraulic anatomy (larger conducting elements toward the base of the vascular system) minimizes gradients in leaf-specific hydraulic conductance with plant height, allowing uniform photosynthesis across the crown of trees. Tip-to-base conduit widening has also been associated with changes in conduit number. However, in bryophytes, despite having representatives with internal water-conducting tissue, conduit widening has been scarcely investigated. Here, we examined the changes in hydroid diameter and number with distance from plant tip in Dawsonia superba and D. polytrichoides, two representatives of the genus containing the tallest extant bryophytes. The position of these moss species on the global scale of conduit size and plant size was consistent with a general scaling among plants with internal water transport. Within plants, patterns of conduit widening and number with distance from plant tip in endohydric mosses were similar to those observed in vascular plants. This study demonstrated that land plants growing upward in the atmosphere show analogous conduit widening of hydraulic structures, suggesting that efficient internal water transport is a convergent adaptation for photosynthesis on land.


Can in vitro studies aid in the development and use of antiseizure therapies? A report of the ILAE/AES Joint Translational Task Force.
In vitro preparations (defined here as cultured cells, brain slices, and isolated whole brains) offer a variety of approaches to modeling various aspects of seizures and epilepsy. Such models are particularly amenable to the application of anti-seizure compounds, and consequently are a valuable tool to screen the mechanisms of epileptiform activity, mode of action of known anti-seizure medications (ASMs), and the potential efficacy of putative new anti-seizure compounds. Despite these applications, all disease models are a simplification of reality and are therefore subject to limitations. In this review, we summarize the main types of in vitro models that can be used in epilepsy research, describing key methodologies as well as notable advantages and disadvantages of each. We argue that a well-designed battery of in vitro models can form an effective and potentially high-throughput screening platform to predict the clinical usefulness of ASMs, and that in vitro models are particularly useful for interrogating mechanisms of ASMs. To conclude, we offer several key recommendations that maximize the potential value of in vitro models in ASM screening. This includes the use of multiple in vitro tests that can complement each other, carefully combined with in vivo studies, the use of tissues from chronically epileptic (rather than naïve wild-type) animals, and the integration of human cell/tissue-derived preparations.


Levator aponeurosis surgery. A retrospective review.
A review of all cases of aponeurotic repair done since 1980 by the author identified 140 patients and 174 eyelids. An acceptable result at 6 weeks was defined as a lid level within 1 mm of the desired level achieved in 74% of lids with acquired ptosis. The amount of ptosis present preoperatively was the major factor in our study; levator function and type of anesthesia did not affect the success rate. The most significant finding was a lower long-term success rate due to a drop in lid level clustered at the 2- to 4-month period postoperatively. The authors speculate on the cause for this finding.


Molecular cloning and chromosomal localization of a novel human tracheo-bronchial mucin cDNA containing tandemly repeated sequences of 48 base pairs.
A lambda gt11 cDNA library constructed from human tracheo-bronchial mucosa was screened with a polyclonal antiserum raised to chemically deglycosylated pronase glycopeptides from human bronchial mucins. Out of 20 positives clones, one partial cDNA clone was isolated and allowed to map a novel human tracheo-bronchial mucin gene. It contains 48 nucleotide tandem repeats quite perfectly identical which encodes a protein containing about 50% of hydroxy amino-acids. This clone hybridized to polydisperse messages produced by human tracheo-bronchial and human colonic mucosae. The gene (proposed name MUC 4) from which cDNA is derived maps to chromosome 3.


Effects of beta-glucanase-producing Lactobacillus strains on growth, dry matter and crude protein digestibilities and apparent metabolisable energy in broiler chickens.
The effects of beta-glucanase expressed by transformed Lactobacillus strains on growth performance, apparent digestibilities of dry matter and crude protein, and apparent metabolisable energy were studied. Two hundred and forty 1-d-old chicks (Avian-43) were randomly divided into three dietary treatment groups and fed with the following diets: (i) basal diet (control) (BD); (ii) basal diet with parental Lactobacillus strains (BDP) and (iii) basal diet with transformed Lactobacillus strains (BDT). At 21 d of age, the body weight, body weight gain and feed conversion ratio of the BDT-fed chickens were significantly improved. At 14 and 21 d of age, the proportions of dry matter in the duodenum, jejunum, ileum, caeca and excreta of chickens given the BDT diet were significantly higher than those of chickens given the BD and BDP diets. Apparent metabolisable energy, digestibilities of crude protein and dry matter were also significantly improved (by 3.5, 5.6 and 3.5%, respectively) by the BDT diet. These results showed that the transformed Lactobacillus strains improved digestibility as well as enhanced the growth performance of chickens.


Malignant phyllodes tumor metastatic to bilateral ovaries: A Krukenberg-like presentation.
Phyllodes tumor (PT) is an extremely rare tumor of the breast of mixed mesenchymal and epithelial origin. It may pursue a benign or malignant evolution with distant metastases in the latter case in 3-12% of patients. The most common sites of metastases are the lungs and bones. Although theoretically any organ may have metastasis, it is extremely rare that a PT will metastasize to the bilateral ovaries and present as Krukenberg tumor. Herein, we report such a case.


The comparison study of bioactivity between composites containing synthetic non-substituted and carbonate-substituted hydroxyapatite.
Apatite forming ability of hydroxyapatite (HAP) and carbonate hydroxyapatite (CHAP) containing composites was compared. Two composite materials, intended for filling bone defects, were made of polysaccharide polymer and one of two types of hydroxyapatite. The bioactivity of the composites was evaluated in vitro by soaking in a simulated body fluid (SBF), and the formation of the apatite layer was determined by scanning electron microscopy with energy-dispersive spectrometer and Raman spectroscopy. The results showed that both the composites induced the formation of apatite layer on their surface after soaking in SBF. In addition, the sample weight changes and the ion concentration of the SBF were scrutinized. The results showed the weight increase for both materials after SBF treatment, higher weight gain and higher uptake of calcium ions by HAP containing scaffolds. SBF solution analysis indicated loss of calcium and phosphorus ions during experiment. All these results indicate apatite forming ability of both biomaterials and suggest comparable bioactive properties of composite containing pure hydroxyapatite and carbonate-substituted one.


Toxic essential oils. Part II: chemical, toxicological, pharmacological and microbiological profiles of Artemisia annua L. volatiles.
Botanical drugs based on Artemisia annua L. (Asteraceae) are important in the treatment of malaria. Alongside with artemisinin, this aromatic species produces high and variable amounts of other chemicals that have mostly unknown biological/pharmacological activities. Herein, we have studied the toxicological/pharmacological profile of volatile constituents of a Serbian population of A. annua. Fifty-eight components were identified, among them, artemisia ketone (35.7%), α-pinene (16.5%) and 1,8-cineole (5.5%) were the most abundant ones. Significant variability of A. annua volatile profile was confirmed by means of agglomerative hierarchical cluster analysis indicating the existence of several different A. annua chemotypes. In an attempt to connect the chemical profile of A. annua oil with its biological/toxicological effects, we have evaluated in vivo and/or in vitro toxicity (including hepato- and nephrotoxicity/protection), antinociceptive, antioxidant (DPPH, ABTS and superoxide radical scavenging activity assays), enzyme inhibiting (protein kinase A and α-amylase) and antimicrobial potential of A. annua oil and of its constituents. Our results revealed that the beneficial properties of A. annua botanical drugs are not limited only to their antimalarial properties. Taking into account its relatively low toxicity, the usage of A. annua volatiles (at least of the herein studied population) does not represent a health risk.


On the fly swapping algorithm for ordering of degrees of freedom in density matrix renormalization group.
Density matrix renormalization group (DMRG) and its time-dependent variants have found widespread applications in quantum chemistry, includingab initioelectronic structure of complex bio-molecules, spectroscopy for molecular aggregates, and charge transport in bulk organic semiconductors. The underlying wavefunction ansatz for DMRG, matrix product state (MPS), requires mapping degrees of freedom (DOF) into a one-dimensional topology. DOF ordering becomes a crucial factor for DMRG accuracy. In this work, we propose swapping neighboring DOFs during the DMRG sweeps for DOF ordering, which we term 'on the fly swapping' (OFS) algorithm. We show that OFS is universal for both static and time-dependent DMRG with minimum computational overhead. Examples are given for one dimensional antiferromagnetic Heisenberg model,ab initioelectronic structure of N2molecule, and the S1/S2internal conversion dynamics of pyrazine molecule. It is found that OFS can indeed improve accuracy by finding better DOF ordering in all cases.


Contrasting roles of E2F2 and E2F3 in endothelial cell growth and ischemic angiogenesis.
The growth of new blood vessels after ischemic injury requires endothelial cells (ECs) to divide and proliferate, and the E2F transcription factors are key regulators of the genes responsible for cell-cycle progression; however, the specific roles of individual E2Fs in ECs are largely unknown. To determine the roles of E2F2 and E2F3 in EC proliferation and the angiogenic response to ischemic injury, hind-limb ischemia was surgically induced in E2F2(-/-) mice, endothelial-specific E2F3-knockout (EndoE2F3(∆/∆)) mice, and their littermates with wild-type E2F2 and E2F3 expression. Two weeks later, Laser-Doppler perfusion measurements, capillary density, and endothelial proliferation were significantly greater in E2F2(-/-) mice and significantly lower in EndoE2F3(∆/∆) mice than in their littermates, and EndoE2F3(∆/∆) mice also developed toe and limb necrosis. The loss of E2F2 expression was associated with increases in the proliferation and G1/S-phase gene expression of isolated ECs, while the loss of E2F3 expression led to declines in these parameters. Thus E2F2 impairs, and endothelial E2F3 promotes, the angiogenic response to peripheral ischemic injury through corresponding changes in EC cell-cycle progression.


Ionic titanocene complexes: a new type of antitumor agent.
Five ionic cyclopentadienyltitanium (IV) derivatives were investigated for their activity against fluid Ehrlich ascites tumor. Four compounds were built up by the intact bis(cyclopentadienyl)titanium(IV) ("titanocene") unit, forming the cationic moiety together with two covalently bound ligands, with certain anions being bonded via electrostatic forces: the acetonitrile complex [(C5H5)2TiCl(NCCH3)]+[FeC14]- (I), the 2'2'-bipyridyl derivative [(C5H5)2Ti(bipy)]2+[CF3SO3]2 (II), the o-phenanthroline complex [(C5H5)2Ti(phen)]2+[CF3SO3]2 (III), and the N-methyl-o-aminothiophenolate derivative [(C5H5)2Ti[o-S(NACH3)C6H4]]+I- (IV). Another ionic cyclopentadienyltitanium derivative investigated was the five-coordinate bis(dithiolene) chelate (C5H5)Ti(1,2,4-S2C6H3CH3)2]-N(C2H5)4)+ (V), the cyclopentadienyltitanium moiety representing the anionic part of the complex salt. All complexes were ionic, salt-like compounds, distinguished by good water solubility. Whereas complexes I, III, and V, given at optimal dose levels, effected maximal cure rates of only 70%-80%, all animals were cured after receiving complexes II and IV at dose ranges of 200-220 and 240-300 mg/kg, respectively. The antitumor activity of complex I was confirmed against solid experimental tumor systems B16 melanoma, colon 38 carcinoma, and Lewis lung carcinosarcoma. Because of their improved solubility in water and pronounced antitumor activity (especially that of II and IV against fluid Ehrlich ascites tumor), ionic cyclopentadienyl titanium complexes are considered to be an interesting new type of antitumor agent.


Infection and utilization rates of bone allografts in a hospital-based musculoskeletal tissue bank in north India.
The bone bank unit of interest in this article was established in January 2018, in a tertiary care teaching institute of north India. Aim of this article is to describe the sources of allografts obtained, discard rates of allografts and infection rates in the recipients after use. All the relevant details of donors and recipients were maintained, and donors were screened for standard inclusion and exclusion criteria before obtaining the grafts. Aerobic culture was performed before storage and just prior to use. Samples with incomplete documentation, incomplete donor screening or positive cultures were discarded. Data on surgical site infection in recipients was collected from hospital records retrospectively. Initially ELISA based serological tests were used for screening. Donor has to undergo these tests again after 6 months to account for the window period of proliferation of viruses. Nucleic acid amplification tests (NAAT) for these viral agents were introduced in the hospital in May 2018. Allografts from a total of 196 donors were obtained in the bone bank over 2 years. Major source of bone was femoral heads harvested during total hip arthroplasty or hemi-arthroplasty. 44(22.4%) grafts had to be discarded. 95 allografts were used in 88 patients during this time. Most common indication for use was surgery for bone tumors (40%), followed by complex primary or revision arthroplasty (30.5%). Three (3.4%) recipients developed deep infection postoperatively. Frozen allograft bone from hospital based bone banks is a reliable source of allografts. When meticulous precautions for sterility are followed, risk of infection is low. Monitoring of such bone banks should fall within a framework of the local legislature. Incomplete documentation is the major reason for wastage of the samples obtained. NAAT may be useful in screening of donors, as it reduces the wastage and the holding time of the allografts.


Prevalence of and factors associated with advanced HIV disease among newly diagnosed people living with HIV in Guangdong Province, China.
A high proportion of people living with HIV (PLHIV) present for care with advanced HIV disease (AHD), which is detrimental to "90-90-90" targets to end AIDS by 2030. This study aimed to explore the prevalence of and factors related to AHD among newly diagnosed PLHIV in Guangdong Province, China. Newly diagnosed PLHIV were recruited from six cities in Guangdong Province from May 2018 to June 2019. AHD was defined as an initial CD4 count <200 cells/µL or an AIDS-defining event within one month of HIV diagnosis. Data from a questionnaire and the national HIV surveillance system were used to explore the potential factors related AHD. A total of 400 of 997 newly diagnosed PLHIV were defined as having AHD with a proportion of 40.1%. After adjusting for statistically significant variables in univariate analysis, multivariable logistic regressions showed that individuals aged 30 to 39 years (adjusted odds ratio (aOR) = 1.77, 95% confidence interval (CI): 1.13 to 2.79) and ≥50 years (aOR = 1.98, 95% CI: 1.15 to 3.43) were at a higher risk of AHD than those aged 18 to 29 years. Participants diagnosed by voluntary counselling and testing (VCT) clinics were less likely to have AHD (aOR = 0.67, 95% CI: 0.48 to 0.94) than those diagnosed at medical facilities. Participants who had ever considered HIV testing (aOR = 0.66, 95% CI: 0.45 to 0.98) and who had high social support (aOR = 0.73, 95% CI: 0.55 to 0.97) were at a lower risk of AHD, whereas participants who had HIV-related symptoms within one year before diagnosis were at a higher risk of AHD (aOR = 2.09, 95% CI: 1.58 to 2.77). The most frequent reason for active HIV testing was "feeling sick" (42.4%, 255/601), and the main reason for never considering HIV testing was "never thinking of getting HIV" (74.0%, 542/732). Low-risk perception and a lack of awareness of HIV-related symptoms resulted in a high proportion of AHD in Guangdong Province, especially among the elderly, those diagnosed at medical facilities and those with low social support. Strengthening AIDS education and training programmes to scale up HIV testing through provider-initiated testing and counselling in medical facilities and VCT could facilitate early HIV diagnosis.


Perioperative blood loss reduction using a sterile exsanguination tourniquet for orthopedic femoral-related surgeries in children: a randomized controlled study.
The sterile exsanguination tourniquet (SET) could be an alternative for providing bloodless surgeries in orthopedic femoral-related surgeries in pediatric patients where the standard pneumatic tourniquet would not be feasible. This randomized-controlled study aimed to evaluate the efficacy of SET in decreasing total perioperative blood loss and blood transfusion. We conducted an unplanned interim analysis of data from a randomized-controlled trial. At the time of the analysis, 31 pediatric patients had been randomly assigned to undergo surgery with the SET application (the SET group, 15 patients) and without the SET application (the control group, 16 patients). An intention-to-treat analysis was performed to evaluate the total perioperative blood loss, postoperative blood transfusion, estimated intraoperative blood loss, total drainage volume, postoperative hemoglobin level, and operative time according to the significance level adjusted for multiplicity (p < 0.029). There was a borderline statistically significant lower body weight-adjusted TBL in the SET group (SET = 14.1 (7.7, 16.9) ml/kg vs. control 18.3 (14.8, 37.2) ml/kg, p-value = 0.027). The body weight-adjusted transfusion volume was statistically significantly greater in the control group (SET = 0.0 (0.0, 0.0) ml/kg vs. control = 2.1 (0.0, 9.7) ml/kg, p = 0.017). Body weight-adjusted estimated intraoperative blood loss was significantly lower in the SET group (SET = 0.8 (0.2, 3.5) ml/kg vs. control = 5.6 (3.4, 21.5) ml/kg, p < 0.001). In addition, the operative time was lower in the SET group with borderline statistical significance (SET = 105 (85.0, 125.0) vs. control = 130 (101.3, 167.5), p = 0.039). Utilization of a sterile exsanguination tourniquet (SET) significantly reduced an estimated intraoperative blood loss while preventing the need for blood transfusion after pediatric orthopedic femoral-related surgeries. Trial registration TCTR20220412003.


Pathology of the olfactory nerve.
The olfactory system and especially the olfactory bulb (OB) as the first relay in the olfactory system represent highly plastic structures. For example, OB volume partly reflects the degree of afferent neural activity. Research indicates that smell deficits leading to a reduced sensory input result in structural changes at the level of the OB. Reduced OB volumes also may be considered characteristic of parosmia. Apart from discussing the clinical implications of these findings, the radiologic basics for assessment of olfactory-eloquent structures are addressed in detail.


Preparation of zein/chitosan/eugenol/curcumin active films for blueberry preservation.
This study aimed to develop zein/chitosan-based films formulated with curcumin and eugenol to improve the quality of postharvest blueberries. First, the film-forming solutions were characterized (rheological property and water distribution), and the mechanical, structural properties and bioactivity of active films fabricated by casting were evaluated. Further, the active film was coated with blueberry stored at 4 °C, and physicochemical properties (weight loss, hardness, microbial counts, and appearance changes) were measured. The film-forming solutions exhibited non-Newtonian behavior. The incorporation of curcumin and eugenol improved the tensile strength and elongation at the break of films, reaching 17.86 MPa and 92.80 %, respectively. The antioxidant capacity was enhanced, and DPPH and ABTS radical scavenging rates were up to 90.60 ± 0.06 % and 86.34 ± 0.39 %, respectively. Meanwhile, the prepared active films possessed good anti-UV and sensitive pH-response color-shifting ability. Compared to the uncoated blueberry, blueberry coated with zein/chitosan/curcumin/eugenol showed lower weight loss and higher hardness, indicating that the prepared active films played a vital role in delaying the deterioration of blueberry and extending its shelf life. Overall, the zein-chitosan incorporated with curcumin and eugenol films could be a promising candidate to prolong the shelf life of food products due to their excellent bioactive capacity.


Interferon production by cells infected with subacute sclerosing panencephalitis (SSPE) virus or measles virus.
Subacute sclerosing panencephalitis (SSPE) is a rare progressive neurodegenerative encephalitis caused by some variants of measles virus (MV). The structure of SSPE virus in the brains of SSPE patients is different from that of MV. The difference in interferon (IFN) production between cells infected with SSPE virus and those infected with MV remains unclear. We measured the concentrations of IFN-α, β, γ, and λ1 (interleukin (IL)-29) from MV- or SSPE virus-infected B95a cells (a marmoset B-lymphoblastoid cell line). SSPE virus-infected B95a cells produced significantly higher levels of IFN-α and λ1 than did MV-infected or mock-infected cells. Our results suggest that SSPE virus and MV induce different IFN production profiles.


Variability of the blood:breath alcohol ratio in vivo.
The blood:breath alcohol ratio, commonly used to translate the result of breath alcohol analysis into the co-existing blood alcohol concentration, varies from person to person and within one person over time.


Salmonella enterica serovar Typhimurium genetic variants isolated after lethal treatment with Thymbra capitata essential oil (TCO) showed increased resistance to TCO in milk.
The high prevalence of Salmonella enterica in milk poses a risk of considerable concern in the preservation of certain dairy products, mainly those elaborated from raw milk. Essential oils (EOs) have been proposed as a promising food preservative for such products due to their strong antimicrobial properties. Additionally, these natural antimicrobials have been shown to be effective against multi-drug resistant strains. They can thus also be utilized to prevent the dissemination of antimicrobial resistances (AMR). However, recent evidence of the development of bacterial resistance under EO treatments may call their use into question. This study sought to assess the emergence of antimicrobial resistant genetic variants of S. enterica serovar Typhimurium from survivors after cyclic exposure to lethal doses (>5 log10 cycles of inactivation) of Thymbra capitata EO (TCO), in order to evaluate the impact that it could have on milk preservation, to ascertain whether cross-resistance to antibiotics occurs, and to identify the genomic changes responsible for their phenotype. Isolated strains by TCO (SeTCO) showed a two-fold increase in minimum inhibitory and bactericide concentrations (MIC and MBC) of TCO compared to Salmonella enterica serovar Typhimurium wild-type strain (SeWT) in laboratory growth medium, as well as a greater adaptation and growth rate in the presence of the EOs and a higher survival to TCO treatments in buffers of pH 4.0 and 7.0. The increased resistance of SeTCO was confirmed in skimmed milk: 300 μL/L TCO reduced only 1 log10 cycle of SeTCO population, whereas it inactivated more than 5 log10 cycles in SeWT. Moreover, SeTCO showed an increased cross-resistance against aminoglycosides, quinolones and tetracyclines. Whole genome sequencing revealed 5 mutations in SeTCO: 2 in genes involved in O-antigens synthesis (rfbV and rfbX), 2 in genes related to adaptation to the growing medium (trkA and glpK), and 1 in a redox-sensitive transcriptional regulator (soxR). The phenotypic characterization of a constructed SeWT strain with mutant soxRSeTCO demonstrated that the mutation of soxR was the main cause of the increased resistance and tolerance observed in SeTCO against TCO and antibiotics. The emergence of resistant strains against EOs might jeopardize their use as food preservatives. Further studies will thus be required to determine under which conditions such resistant strains might occur, and to assess the food risk they may pose, as well as to ascertain their impact on the spread of AMR.


Increasing Blood Pressure by Greater Splanchnic Nerve Stimulation: a Feasibility Study.
The splanchnic vascular compartment is the major reservoir for intravascular blood volume, and dysregulation of the compartment was implicated in a series of cardiovascular conditions. We explored feasibility and effectiveness of an implantable cuff system on the greater splanchnic nerve (GSN) in healthy canines for short- and long-term neuromodulation to affect the circulation. Five mongrel hounds underwent minimally invasive right-sided unilateral GSN cuff placement. All animals underwent same day GSN stimulation and repeat stimulation at 9-30 days. Stimulation parameter optimization was conducted both acutely and chronically. Parameters ranged from 1-250 Hz, 0.25 mA-35 mA, 0.1-0.5 ms, and 30-s pulse duration. Two animals were survived for 9 days and 3 animals for 30 days. Stimulation of the right GSN increased mean arterial blood pressure by 36.9 mmHg ± 13.4 (p < 0.0001), central venous pressure by 6.9 mmHg ± 1.7 (p < 0.0001), and mean pulmonary arterial pressure by 6.3 mmHg ± 2.0 (p < 0.0001). Peak effects were observed within 30 s, and magnitude of effects was comparable between stimulation cycles (p = 0.4). Stimulation-induced changes in hemodynamics were independent of afferent nerve fibers (pain response) or the adrenal gland. Necropsy showed no evidence of nerve damage on histologic studies up to 30 days after implantation. GSN stimulation via an implanted nerve cuff provided a reproducible and rapid method to increase arterial, central venous, and pulmonary arterial pressures. The neuromodulation cuff was well tolerated and elicited a response up to 30 days after implantation. The clinical application of GSN stimulation as a tool to change central and peripheral cardiovascular hemodynamics needs to be explored.


The negotiation of the sick role: general practitioners' classification of patients with medically unexplained symptoms.
In encounters between general practitioners (GPs) and patients with medically unexplained symptoms (MUS), the negotiation of the sick role is a social process. In this process, GPs not only use traditional biomedical diagnostic tools but also rely on their own opinions and evaluations of a patient's particular circumstances in deciding whether that patient is legitimately sick. The doctor is thus a gatekeeper of legitimacy. This article presents results from a qualitative interview study conducted in Denmark with GPs concerning their approach to patients with MUS. We employ a symbolic interaction approach that pays special attention to the external validation of the sick role, making GPs' accounts of such patients particularly relevant. One of the article's main findings is that GPs' criteria for judging the legitimacy of claims by those patients that present with MUS are influenced by the extent to which GPs are able to constitute these patients as people with social problems and problematic personality traits.


Computer based information systems in primary health care--why?
The delivery of health care is information based. A host of computer-based information systems have been developed and implemented in the health care environment. The mere availability of the computer as a tool for information handling should, in itself, not be the cause for developing computer-based information systems. The earlier assumption of a cost-benefit impact with the development of such systems has not been effectively shown in all cases, and as seen in a report by van Bemmel, the recent trend is to apply other criteria in systems evaluation. Information is essential in health care related decision making. The properties of information are described to present a case for a computer-based information system to support primary health care delivery.


Natural selection in neoplastic progression of Barrett's esophagus.
Neoplasms progress to cancer through a process of natural selection. The rate of evolution, and thus progression is determined by three parameters: mutation rate, population size of the evolving neoplastic cells, and intensity of selection or rate of clonal expansion. All three parameters are reviewed in the context of Barrett's esophagus, a pre-malignant neoplasm. Although Barrett's esophagus is an ideal model for the study of neoplastic clonal evolution, similar studies may be carried out in a wide variety of human neoplasms. Evolutionary analyses provide insights for clinical management, including rates of progression to cancer and emergence of resistance to interventions.


Quantitative dynamic contrast-enhanced ultrasound to predict intrahepatic recurrence of hepatocellular carcinoma after radiofrequency ablation: a cohort study.
Quantitative dynamic contrast-enhanced ultrasonography (CEUS) reflects tumor blood perfusion. There are very few studies on the relationship between intrahepatic recurrence of hepatocellular carcinoma (HCC) and tumor perfusion. We investigated the correlation of dynamic CEUS parameters with intrahepatic recurrence after radiofrequency ablation (RFA). This retrospective study enrolled 125 native HCC patients who underwent RFA between September 2017 and January 2019 with curative intent. Pre-ablation quantitative dynamic CEUS was performed. CEUS parameters were extracted from time-intensity curves. The correlation of CEUS parameters with intrahepatic recurrence was investigated. The mean follow-up time was 21.6 ± 7.9 months. The recurrence rate was 33.6%. Univariate and multivariate analyses revealed that tumor peak intensity (PI) was a significant independent risk factor for intrahepatic recurrence after RFA (hazard ratio (HR), 0.3; 95% CI, 0.1-0.9). A PI of 58.8% (area under curve, 0.72; 95% CI, 0.63-0.81) was considered as the optimal cutoff level to predict the intrahepatic recurrence of HCC in patients after RFA. The recurrence-free survival rate in patients with a PI > 58.8% was 94.4% at 1 year and 77.8% at 2 years. Subgroup analysis showed that the HR of time to peak (TTP) in intrahepatic recurrence was 1.1417 (95% CI, 0.9748-1.1436; p = 0.1973) in the patient group with tumor diameter > 31 mm. CEUS is commonly used in HCC patients who undergo RFA. The CEUS parameters PI and TTP are associated with intrahepatic recurrence after RFA, and can thus be used to identify patients at risk for intrahepatic recurrence.


Evidence of histidine phosphorylation in isocitrate lyase from Escherichia coli.
Escherichia coli isocitrate lyase (EC 18.104.22.168.) can be phosphorylated in vitro by an ATP-dependent reaction. The enzyme becomes phosphorylated by an endogenous kinase when partially purified sonic extracts are incubated with [gamma-32P]ATP. Treatment of isocitrate lyase with diethyl pyrocarbonate, a histidine-modifying reagent, blocked incorporation of [32P]phosphate from [gamma-32P]ATP. The isoelectric point of the enzyme was altered by treatment with phosphoramidate, a histidine phosphorylating agent, which suggests that isocitrate lyase can be phosphorylated at a histidine residue(s). Immunoprecipitated 32P-labeled isocitrate lyase was subjected to alkaline hydrolysis, mixed with chemically synthesized phosphohistidine standards, and analyzed by anion exchange chromatography. Characterization of the phosphoamino acid was based on the demonstration that the 32P-labeled product from alkali-hydrolyzed isocitrate lyase comigrated with synthetic 1-phosphohistidine. In addition, loss of catalytic activity after treatment with potato acid phosphatase indicates that catalytically active isocitrate lyase is the phosphorylated form of the enzyme.


Should silicone prostheses be considered for specimen banking? A pilot study into their use for human biomonitoring.
Persistent organic pollutant (POP) biomonitoring in humans is challenging and generally carried out using blood, breast milk or adipose tissue, with concentrations normalised to the lipid content of the sample matrix. The goal of this cross-sectional pilot study was to evaluate the validity and feasibility of explanted silicone prostheses as a matrix for persistent organic pollutant biomonitoring in humans. We postulate that pollutant concentrations in silicone prostheses inserted in the body will equilibrate with that in the body over time and provide a measure of the overall body burden. This study included silicone prostheses from 22 female patients of the Colosseum clinic (Oslo, Norway) collected between September 2010 and April 2012. Absorption of chlorinated and brominated POPs into silicone prostheses during implantation was observed. Relative levels of the different contaminants measured in prostheses were in agreement with those from serum and breast milk analyses from the general Norwegian population. The comparison of serum and breast milk-based literature data with prosthesis concentrations transposed into lipid-normalised concentrations supports the validity of the prosthesis measurements. The median of relative percent differences between measurements with replicate silicone prostheses from 11 patients was below 30%. Observed increases in prosthesis concentrations with patients' age were found to be very similar to literature data from studies of the Norwegian population. Silicone prostheses therefore represent a promising matrix for the biomonitoring of nonpolar and non-ionic pollutants in humans. Sample accessibility and body burden representativeness of the silicone prostheses suggest that specimen banking should be initiated.


Genetic variation in vitamin D-related genes and risk of breast cancer among women of European and East Asian descent.
Studies of vitamin D-related genetic variants and breast cancer have been inconsistent. This study aimed to investigate associations of vitamin D-related polymorphisms and breast cancer risk among European and East Asian women and potential interactions with menopausal status and breast tumour subtypes. Data from a case-control study of breast cancer (1037 cases and 1050 controls) were used to assess relationships between 21 polymorphisms in two vitamin D-related genes (GC and VDR) and breast cancer risk. Odds ratios were calculated in stratified analyses of European and East Asian women, using logistic regression in an additive genetic model. An interaction term was used to explore modification by menopausal status. Polytomous regression was used to assess heterogeneity by breast tumour subtype. False discovery rate adjustments were conducted to account for multiple testing. No association was observed between GC or VDR polymorphisms and breast cancer risk. Modification of these relationships by menopausal status was observed for select polymorphisms in both Europeans (VDR rs4328262 and rs11168292) and East Asians (GC rs7041 and VDR rs11168287). Heterogeneity by tumour subtype was seen for three VDR polymorphisms (rs1544410, rs7967152 and rs2239186) among Europeans, in which associations with ER-/PR-/HER2+ tumours, but not with other subtypes, were observed. In conclusion, associations between vitamin D-related genetic variants and breast cancer were not observed overall, although the relationships between vitamin D pathway polymorphisms and breast cancer may be modified by menopausal status and breast tumour subtype.