Ginsenoside Rg1 Alleviates Ulcerative Colitis in Obese Mice by Regulating the Gut Microbiota-Lipid Metabolism-Th1/Th2/Th17 Cells Axis.
Ginsenoside Rg1 (G-Rg1) has various pharmacological properties including antiobesity, immunomodulatory, and anti-inflammatory effects. This study aimed to explore the therapeutic effects and underlying mechanisms of G-Rg1 on colitis complicated by obesity. The results indicate that G-Rg1 effectively alleviates colitis in obese mice and improves serum lipid levels and liver function. Importantly, G-Rg1 improved the composition of gut microbiota in obese mice with colitis, with increases in alpha diversity indexes Sobs, Ace, and Chao, a significant down-regulation of the relative abundance of Romboutsia, and a significant up-regulation of Rikenellaceae_RC9_gut_group, Lachnospiraceae_NK4A136_group, Enterorhabdus, Desulfovibrio, and Alistipes. Meanwhile, G-Rg1 improved lipid metabolism in the colonic contents of obese mice with colitis. Additionally, G-Rg1 significantly reduced the percentages of helper T (Th)1, Th17, central memory T (TCM), and effector memory T (TEM) cells in obese mice with colitis while significantly increasing Naïve T and Th2 cells. In conclusion, G-Rg1 could be a promising therapeutic option for alleviating obesity complicated by colitis through regulation of the gut microbiota and lipid metabolism as well as Th1/Th2/Th17 cell differentiation.


Two-dimensional NMR and all-atom molecular dynamics of cytochrome P450 CYP119 reveal hidden conformational substates.
Cytochrome P450 enzymes are versatile catalysts involved in a wide variety of biological processes from hormonal regulation and antibiotic synthesis to drug metabolism. A hallmark of their versatility is their promiscuous nature, allowing them to recognize a wide variety of chemically diverse substrates. However, the molecular details of this promiscuity have remained elusive. Here, we have utilized two-dimensional heteronuclear single quantum coherence NMR spectroscopy to examine a series of mutants site-specific labeled with the unnatural amino acid, [(13)C]p-methoxyphenylalanine, in conjunction with all-atom molecular dynamics simulations to examine substrate and inhibitor binding to CYP119, a P450 from Sulfolobus acidocaldarius. The results suggest that tight binding hydrophobic ligands tend to lock the enzyme into a single conformational substate, whereas weak binding low affinity ligands bind loosely in the active site, resulting in a distribution of localized conformers. Furthermore, the molecular dynamics simulations suggest that the ligand-free enzyme samples ligand-bound conformations of the enzyme and, therefore, that ligand binding may proceed largely through a process of conformational selection rather than induced fit.


Hairdressers as a source of social support: A qualitative study on client disclosures from Australian hairdressers' perspectives.
Hairdressing is one of the few non-health professional occupations where workers physically interact with clients, who are often comfortable in confiding private information to them. Despite this, relatively little is known about how hairdressers understand and experience this role as informal confidants. This study aimed to address this using an in-depth qualitative approach to capture what hairdressers are hearing from clients, and how hairdressers respond and feel about supporting clients. Participant led interviews with prompt questions were conducted in South Australia in 2019, transcribed and analysed using Thematic Analysis. Results indicate clients disclose information about family, health, identity, mental health and women's health. Important themes generated in relation to the role of hairdressers included: their need to have a client focus; blurring role boundaries; behaving like a therapist and, providing a place of safety and advice, while also maintaining confidentiality. Hairdressers reported feeling undervalued, emotionally drained, and in need of support in their role, but conversely, also reported having good job satisfaction. Further research is needed to examine whether hairdresser-client interactions may promote better mental and physical health outcomes for clients due to psychological buffering arising from the social support provided by hairdressers, as well as the impact on hairdressers and implications for their training.


A Limulus antilipopolysaccharide factor-derived peptide exhibits a new immunological activity with potential applicability in infectious diseases.
Previous studies have shown that cyclic peptides corresponding to residues 35 to 52 of the Limulus antilipopolysaccharide (anti-LPS) factor (LALF) bind and neutralize LPS-mediated in vitro and in vivo activities. Therapeutic approaches based on agents which bind and neutralize LPS activities are particularly attractive because these substances directly block the primary stimulus for the entire proinflammatory cytokine cascade. Here we describe new activities of the LALF(31-52) peptide, other than its LPS binding ability. Surprisingly, supernatants from human mononuclear cells stimulated with the LALF peptide are able to induce in vitro antiviral effects on the Hep-2 cell line mediated by gamma interferon (IFN-gamma) and IFN-alpha. Analysis of the effect of LALF(31-52) on tumor necrosis factor (TNF) and nitric oxide (NO) production by LPS-stimulated peritoneal macrophages revealed that a pretreatment with the peptide decreased LPS-induced TNF production but did not affect NO generation. This indicates that the LALF peptide modifies the LPS-induced response. In a model in mice with peritoneal fulminating sepsis, LALF(31-52) protected the mice when administered prophylactically, and this effect is related to reduced systemic TNF-alpha levels. This study demonstrates, for the first time, the anti-inflammatory properties of the LALF-derived peptide. These properties widen the spectrum of the therapeutic potential for this LALF-derived peptide and the molecules derived from it. These agents may be useful in the prophylaxis and therapy of viral and bacterial infectious diseases, as well as for septic shock.


Identification of Causes of the Occupational Stress for Health Providers at Different Levels of Health Care.
To identify and compare the stressors in the work environment experienced by employees in primary health care and secondary health care, amongphysiciansand nurses. The survey was conducted to identify types of stressors by assessing health care workers employed in the primary and secondary health care services of the Public Institution, the Health Centre of the Sarajevo Canton, using a questionnaire about stress in the workplace. Among all study participants stressors connected to the organization of work, finance and communication were found to affect their mental health most strongly. The results show a significant difference between primary and secondary health care in experience of stressors related to the organization of work, communication, and stressors related to the emotional and physical risks. Primary health care physicians report a significantly higher experience of stress and impact on mental health compared with other physicians related to emotional difficulties when working in the field of palliative care. Our results also indicate a significant difference between primary and secondary health providers in experiencing stressors related to the organization of work, such as: on-call duty shifts, an inadequate working environment and in the assessment of administrative work overload. The survey identified the most intense stressors for doctors and nurses at primary and secondary levels of health care services. The results of the study indicate that doctors and nurses have a different hierarchy of stressors, as well as subjects at Primary and Secondary Health Care. The results of the study indicate that subjects et Primary Health Care perceive more stressful organizational, emotional and communicational problems.


Effect of seasonal ragweed exposure on immunoglobulin E antiragweed antibodies in cultures of peripheral mononuclear cells, plasma and nasal secretions.
To determine the effects of seasonal ragweed exposure on in vitro IgE antibody production, the antiragweed IgE antibody content of 7-day cultures of peripheral blood mononuclear cells was assessed in samples obtained before, during and after the ragweed season, in 4 ragweed-allergic and 11 nonallergic individuals. The levels of ragweed-specific IgE and total IgE in unstimulated culture supernatants were measured by solid phase radioimmunoassays. Antiragweed IgE antibody was apparently detectable in the 7-day culture supernatants of both allergic and nonallergic individuals, and the antibody concentrations increased after natural ragweed exposure. However, the bindable counts of IgE antibody in supernatants from allergic patients were inhibited by soluble ragweed antigen, while those from nonallergics were not inhibitable, suggesting that supernatants from nonallergics did not contain true ragweed-specific IgE antibody. Because, for most subjects, the quantity of IgE in culture supernatants from 7-day cultures of living cells was the same as that derived from cells which were either frozen and thawed prior to culture, or briefly exposed to pH 3.7 buffer, the majority of IgE antibody in supernatants from allergic subjects, and of total IgE in supernatants from all subjects, was apparently not newly synthesized, but was preformed, and appeared to be largely derived from the cell surface. IgE antiragweed antibody was detected in the nasal secretions and plasma of allergic, but not nonallergic individuals, and the levels of antibody in allergic subjects increased after seasonal exposure. The fraction of total IgE protein accounted for by IgE antiragweed antibody in nasal secretions collected postseasonally was higher than that in both culture supernatants and plasma, suggesting that ragweed-specific IgE antibody is synthesized locally in the nose.


Lessons from assembling a microbial natural product and pre-fractionated extract library in an academic laboratory.
Microbial natural products are specialized metabolites that are sources of many bioactive compounds including antibiotics, antifungals, antiparasitics, anticancer agents, and probes of biology. The assembly of libraries of producers of natural products has traditionally been the province of the pharmaceutical industry. This sector has gathered significant historical collections of bacteria and fungi to identify new drug leads with outstanding outcomes-upwards of 60% of drug scaffolds originate from such libraries. Despite this success, the repeated rediscovery of known compounds and the resultant diminishing chemical novelty contributed to a pivot from this source of bioactive compounds toward more tractable synthetic compounds in the drug industry. The advent of advanced mass spectrometry tools, along with rapid whole genome sequencing and in silico identification of biosynthetic gene clusters that encode the machinery necessary for the synthesis of specialized metabolites, offers the opportunity to revisit microbial natural product libraries with renewed vigor. Assembling a suitable library of microbes and extracts for screening requires the investment of resources and the development of methods that have customarily been the proprietary purview of large pharmaceutical companies. Here, we report a perspective on our efforts to assemble a library of natural product-producing microbes and the establishment of methods to extract and fractionate bioactive compounds using resources available to most academic labs. We validate the library and approach through a series of screens for antimicrobial and cytotoxic agents. This work serves as a blueprint for establishing libraries of microbial natural product producers and bioactive extract fractions suitable for screens of bioactive compounds.


Daily oviposition activity of Aedes aegypti in Orán, Argentina.
The study aimed to determinate the maximum daily peak of Aedes aegypti oviposition in the city of Oran, northwestern Argentina. Biweekly samplings were taken between November 2006 and February 2007 (spring-summer). The city was divided into three areas (north, center, and south) and households were randomly selected. Two ovitraps were placed outdoors in the selected houses. Ovitraps were replaced every four hours, from morning (8 a.m.) to late afternoon (8 p.m.). The largest number of eggs was recorded between 4 p.m. and 8 p.m. (81%). These findings enhance our understanding of the vector and thus its control such as spraying during the hours of peak oviposition activity.


Initial Opioid Prescriptions Predict Continued Narcotic Use: Analysis of 24,594 Reduction Mammaplasty Patients.
The objective of this study was to establish whether a particular perioperative opioid regimen was associated with a higher risk of opioid refilling in the reduction mammaplasty patient population. A retrospective cohort study was conducted on subjects that underwent bilateral reduction mammaplasty with no history of opioid use in the year before surgery. Patients were followed for a period of 4 months after surgery. Multivariable logistic regression was used to establish factors associated with prescription refilling. Predictive probabilities of opioid refilling, given various perioperative opioid regimens, were explored. A total of 24,594 subjects met all criteria for inclusion in the study, 13 percent of whom demonstrated continued opioid use following surgery. The probability of refill in patients receiving 15 to 59 daily morphine milligram equivalents perioperatively was significantly lower than in those receiving less than 15 daily morphine milligram equivalents (e.g., 15 mg of codeine every 4 hours) or more than 60 daily morphine milligram equivalents (e.g., 10 mg of oxycodone every 6 hours). The probability of refilling prescription opioids is increased by nonmodifiable risk factors, such as age. However, modifiable risk factors exist as well, most notably the amount of narcotic prescribed during the perioperative period. Overprescribing was found to be common in the reduction mammaplasty patient population. Limiting outpatient opioid exposure to a range of 15 to 59 daily morphine milligram equivalents (e.g., 5 mg of hydrocodone every 6 hours) during the perisurgical period may reduce the probability that the surgeon will need to prescribe further narcotics. Further studies are needed to validate our findings. Risk, III.


Exploring the optimal electropolymerization strategy for the preparation of solid-phase microextraction fibers using pyrrole-dopamine copolymers.
In-situ electropolymerization of conductive polymers on the surface of stainless-steel substrates is a well-established but promising procedure for the preparation of solid-phase microextraction (SPME) tools. Herein, different electrochemical methods including constant potential (CP), constant potential pulse (CPP), and cyclic voltammetry (CV) were utilized to fabricate SPME fibers by in-situ electropolymerization of pyrrole-dopamine copolymers (PPY/PDA) on the surface of stainless-steel fibers. The coated fibers were characterized and applied for the direct-immersion SPME (DI-SPME) sampling of ultra-trace amounts of plant hormones including abscisic acid (ABA), gibberellic acid (GA3), and indole acetic acid (IAA) in fruit juices, followed by HPLC-UV determination. The results showed that CV electropolymerization is significantly more efficient than the two other methods. The coatings created by the CV method were satisfactorily uniform, adhesive, and durable and exhibited higher extraction performance compared to the CP and CPP procedures. The important experimental variables of the proposed DI-SPME-HPLC method were evaluated and optimized using response surface methodology with a Box-Behnken design. The developed method showed wide-range linearities, spanning from 0.05 to 20μg mL-1 for GA3, and 0.02 to 20μg mL-1 for ABA and IAA. The limits of detection were obtained 0.01μg mL-1 for GA3, and 0.005μg mL-1 for ABA and IAA. The fiber was successfully employed for the simultaneous DI-SPME-HPLC analysis of plant hormones in fruit juice samples.


Inspiratory Muscle Strength in Chagas Cardiomyopathy: A Systematic Scoping Review.
The increase in inflammatory markers associated with persistent chronic fibrosing myocarditis, a characteristic of chronic Chagas disease, can result in a reduction in inspiratory muscle strength (IMS) in Chagas cardiomyopathy (CC). However, literature in this field is still scarce. This review aimed to map and summarize the evidence regarding IMS in patients with CC. The inclusion criteria included reports with adult participants with a CC diagnosis, with or without heart failure (HF). The core concept examined was the maximum inspiratory pressure evaluated in the untrained and trained groups in the pre-training period. The context was open, including but not limited to hospitals and health centers. Two authors independently identified eligible studies and extracted the data. Descriptive synthesis was used as the primary strategy for analyzing the results. Nine studies (five clinical trials, three cross-sectional, and one cohort) were included. The CC classification differed among the studies, with no mention of HF in five and no CC staging specification in six. IMS was assessed using a manovacuometer, and only six studies analyzed and interpreted the data concerning the predicted values. The CC population with HF appeared to have impaired IMS. All studies involved only Brazilian volunteers. In conclusion, randomized clinical trials evaluating IMS and the effects of inspiratory muscle training need to be conducted to better understand the prevalence and risk of inspiratory muscle weakness in the CC population, as well as the effects of training. Such studies should be conducted at different stages of CC in different populations and countries.


Developmenrt of EST-SSR and genomic-SSR markers to assess genetic diversity in Jatropha Curcas L.
Jatropha curcas L. has attracted a great deal of attention worldwide, regarding its potential as a new biodiesel crop. However, the understanding of this crop remains very limited and little genomic research has been done. We used simple sequence repeat (SSR) markers that could be transferred from Manihot esculenta (cassava) to analyze the genetic relationships among 45 accessions of J. curcas from our germplasm collection. In total, 187 out of 419 expressed sequence tag (EST)-SSR and 54 out of 182 genomic (G)-SSR markers from cassava were polymorphic among the J. curcas accessions. The EST-SSR markers comprised 26.20% dinucleotide repeats, 57.75% trinucleotide repeats, 7.49% tetranucleotide repeats, and 8.56% pentanucleotide repeats, whereas the majority of the G-SSR markers were dinucleotide repeats (62.96%). The 187 EST-SSRs resided in genes that are involved mainly in biological and metabolic processes. Thirty-six EST-SSRs and 20 G-SSRs were chosen to analyze the genetic diversity among 45 J. curcas accessions. A total of 183 polymorphic alleles were detected. On the basis of the distribution of these polymorphic alleles, the 45 accessions were classified into six groups, in which the genotype showed a correlation with geographic origin. The estimated mean genetic diversity index was 0.5572, which suggests that our J. curcas germplasm collection has a high level of genetic diversity. This should facilitate subsequent studies on genetic mapping and molecular breeding. We identified 241 novel EST-SSR and G-SSR markers in J. curcas, which should be useful for genetic mapping and quantitative trait loci analysis of important agronomic traits. By using these markers, we found that the intergroup gene diversity of J. curcas was greater than the intragroup diversity, and that the domestication of the species probably occurred partly in America and partly in Hainan, China.


Thalamic Functional Connectivity in Mild Traumatic Brain Injury: Longitudinal Associations With Patient-Reported Outcomes and Neuropsychological Tests.
(1) To examine differences in patient-reported outcomes, neuropsychological tests, and thalamic functional connectivity (FC) between patients with mild traumatic brain injury (mTBI) and individuals without mTBI and (2) to determine longitudinal associations between changes in these measures. Prospective observational case-control study. Academic medical center. A sample (N=24) of 13 patients with mTBI (mean age, 39.3±14.0y; 4 women [31%]) and 11 age- and sex-matched controls without mTBI (mean age, 37.6±13.3y; 4 women [36%]). Not applicable. Resting state FC (3T magnetic resonance imaging scanner) was examined between the thalamus and the default mode network, dorsal attention network, and frontoparietal control network. Patient-reported outcomes included pain (Brief Pain Inventory), depressive symptoms (Patient Health Questionnaire-9), posttraumatic stress disorder ([PTSD] Checklist - Civilian Version), and postconcussive symptoms (Rivermead Post-Concussion Symptoms Questionnaire). Neuropsychological tests included the Delis-Kaplan Executive Function System Tower test, Trails B, and Hotel Task. Assessments occurred at 6 weeks and 4 months after hospitalization in patients with mTBI and at a single visit for controls. Student t tests found increased pain, depressive symptoms, PTSD symptoms, and postconcussive symptoms; decreased performance on Trails B; increased FC between the thalamus and the default mode network; and decreased FC between the thalamus and the dorsal attention network and between the thalamus and the frontoparietal control network in patients with mTBI as compared with controls. The Spearman correlation coefficient indicated that increased FC between the thalamus and the dorsal attention network from baseline to 4 months was associated with decreased pain and postconcussive symptoms (corrected P<.05). Findings suggest that alterations in thalamic FC occur after mTBI, and improvements in pain and postconcussive symptoms are correlated with normalization of thalamic FC over time.


A high-throughput sequencing test for diagnosing inherited bleeding, thrombotic, and platelet disorders.
Inherited bleeding, thrombotic, and platelet disorders (BPDs) are diseases that affect ∼300 individuals per million births. With the exception of hemophilia and von Willebrand disease patients, a molecular analysis for patients with a BPD is often unavailable. Many specialized tests are usually required to reach a putative diagnosis and they are typically performed in a step-wise manner to control costs. This approach causes delays and a conclusive molecular diagnosis is often never reached, which can compromise treatment and impede rapid identification of affected relatives. To address this unmet diagnostic need, we designed a high-throughput sequencing platform targeting 63 genes relevant for BPDs. The platform can call single nucleotide variants, short insertions/deletions, and large copy number variants (though not inversions) which are subjected to automated filtering for diagnostic prioritization, resulting in an average of 5.34 candidate variants per individual. We sequenced 159 and 137 samples, respectively, from cases with and without previously known causal variants. Among the latter group, 61 cases had clinical and laboratory phenotypes indicative of a particular molecular etiology, whereas the remainder had an a priori highly uncertain etiology. All previously detected variants were recapitulated and, when the etiology was suspected but unknown or uncertain, a molecular diagnosis was reached in 56 of 61 and only 8 of 76 cases, respectively. The latter category highlights the need for further research into novel causes of BPDs. The ThromboGenomics platform thus provides an affordable DNA-based test to diagnose patients suspected of having a known inherited BPD.


Insulin resistance in fat cells from insulin-treated type I diabetic individuals.
Fat biopsies from the lower abdominal wall were obtained from 13 insulin-treated type I diabetic subjects and from 12 age-, weight-, and sex-matched control subjects. Insulin binding and the antilipolytic effect of insulin were studied. Insulin binding was significantly reduced in the diabetic subjects (34% reduction at tracer binding, P less than 0.05) due to a decreased number of binding sites. In agreement with this, the dose-response curve for the antilipolytic effect of insulin was shifted to the right in the diabetic subjects. Furthermore, the maximal antilipolytic effect of insulin was also reduced (64%, P less than 0.05). Thus, fat cells from conventionally treated type I diabetic individuals are resistant to insulin. This resistance is due to a combination of a decreased number of insulin binding sites and an unspecified intracellular (postreceptor) defect involving the antilipolytic effect of insulin. These findings are in accord with recent in vivo studies showing that type I diabetic patients are also resistant to the stimulating effect of insulin on glucose disposal.


Long-term GnRH-agonist administration revealed a GnRH-independent mechanism stimulating FSH discharge in humans.
The present study evaluated the FSH and LH episodic discharge in different physiopathological conditions undergoing chronic GnRH-agonist administration. Four girls with true precocious puberty and five postmenopausal women were administered GnRH-agonist (3.73 leuprolide acetate every 4 weeks; Takeda Italia, Rome, Italy) for at least 4 months. Plasma LH and FSH secretory profiles were assessed before and under GnRH-agonist administration (after 21 and 120 days). Pulsatility studies were conducted for 4 h in the girls and for 6 h in postmenopausal women, with blood sampling intervals of 10 min. Pubertal and postmenopausal patients showed the distinct episodic co-secretion of LH and FSH before GnRH-agonist administration; this co-secretion disappeared in both groups after 21 and 120 days of treatment. Moreover, while LH concentrations decreased to almost undetectable levels and LH episodic release disappeared, FSH plasma levels were only partially reduced and FSH episodic secretion was detectable in both groups. In conclusion, this study demonstrated that long-term GnRH-agonist administration blocked LH but not FSH episodic release. These data enforce the hypothesis that FSH episodic discharge might be dependent not only on hypothalamic GnRH, but also on a GnRH-independent stimulatory pathway.


A systematic review of psychological interventions for the treatment of nonadherence to fluid-intake restrictions in people receiving hemodialysis.
Psychological interventions aimed at improving adherence to fluid-intake restrictions in patients receiving hemodialysis have become increasingly common. To the authors' knowledge, this is the first systematic review of the literature examining the impact of these interventions associated with patient interdialytic weight gain (IWG). A systematic search of the literature was performed on EMBASE, MEDLINE, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, and PsychINFO. The search was augmented by manually examining reference lists of reviews and retrieved reports. Study quality was graded according to criteria developed by the authors. Two additional independent researchers separately coded a random sample of studies to avoid bias of rating. Sixteen studies were identified as eligible for inclusion. Relevant information from each included study was extracted and entered into a standardized table. Nearly all studies showed a postintervention decrease in IWG. A number of method weaknesses in the existing literature were identified. Studies investigating psychological interventions aimed at improving adherence to fluid-intake restrictions appear to indicate some success in decreasing IWG. However, confidence regarding the validity of this finding is circumscribed by the prevalent use of investigative designs with inherently high susceptibility to bias. Future studies would benefit from using larger numbers of participants within controlled designs. Clearer description of intervention protocols would foster greater understanding of the contextual appropriateness of different approaches and which treatment components are key to improving adherence to fluid-intake restrictions in patients receiving hemodialysis.


[Women, schizophrenia and oestrogen; neurobiological hypotheses and hormonetherapy studies].
Gender differences play a role in the origin and course of schizophrenia. It has been hypothesised that the gonadal hormone, oestrogen, may possibly perform a protective function in the development of certain forms of schizophrenia. To review neurobiological hypotheses concerning the role of oestrogen in the development and course of schizophrenia. The relevant literature was consulted with the help of PubMed, textbooks and bibliographic references; the search terms used were 'oestrogen', 'schizophrenia', 'gender', 'epigenetics', 'psychosis', 'women' and 'brain'. There were no restrictions with regards to the time-period. Neuro-imaging, animal experiments and hormone-therapy studies showed several effects of oestrogen in the field of epigenetics, morphology of the brain, interaction with neurotransmitters and neuroprotection. Oestrogen is an important link in a complex of factors that clearly play a role in the varying development of schizophrenia in men and women. So far, however, there is insufficient evidence to support the existence of a specific mechanism that would explain why oestrogen may perform a protective function in schizophrenia.


Impact of the viral respiratory season on postoperative outcomes in children undergoing cardiac surgery.
To compare postoperative outcomes in children undergoing cardiac surgery during the viral respiratory season and nonviral season at our institution. This was a retrospective cohort study and secondary matched case-control analysis. The setting was an urban academic tertiary-care children's hospital. The study was comprised of all patients <18 years of age who underwent cardiac surgery at Johns Hopkins Hospital from October 2002 through September 2007. Patients were stratified by season of surgery, complexity of cardiac disease, and presence or absence of viral respiratory infection. Measurements included patient characteristics and postoperative outcomes. The primary outcome was postoperative length of stay (LOS). A total of 744 patients were included in the analysis. There was no difference in baseline characteristics or outcomes, specifically, no difference in postoperative LOS, intensive care unit (ICU) LOS, and mortality, among patients by seasons of surgery. Patients with viral respiratory illness were more likely to have longer postoperative LOS (p < 0.01) and ICU LOS (p < 0.01) compared with matched controls. We identified no difference in postoperative outcomes based on season in patients undergoing cardiac surgery. Children with viral respiratory infection have significantly worse outcomes than matched controls, strengthening the call for universal administration of influenza vaccination and palivizumab to appropriate groups. Preoperative testing for respiratory viruses should be considered during the winter months for children undergoing elective cardiac surgery.


Thermodynamic study of molecularly imprinted polymer used as the stationary phase in high performance liquid chromatography.
Molecularly imprinted polymer (MIP) and non-imprinted polymer (NIP) on the base of methacrylic acid prepared by a bulk polymerization were used as stationary phases for the HPLC analysis. The thermodynamic processes were carried out to investigate the temperature effects during sorption processes of potential local anaesthetics - morpholinoethyl esters of alkoxy-substituted phenylcarbamic acid (MEsP), local anaesthetic - diperodon, flavonoid - quercetin in methanol, acetonitrile and toluene (porogen) as mobile phases. Mobile phases and corresponding solvents were selected according to the solubility of each analyte. The template was chosen from the set of homologous of MEsP - 2-(morpholin-4-yl)ethyl (2-methoxyphenyl)carbamate. Values of retention factors were measured over the temperature range of 20-60°C. There were determined van't Hoff curves - dependences between logarithms of the retention factors (lnk) and the inverse value of the temperature (1/T). Observed graphs were linear directly indicating that there were no changes of interaction mechanisms in the studied range of temperature. Selectivities (evaluated by the separation factors, α) and sorption selectivities (evaluated by the imprinting factors, IFs) of the MIP and the NIP toward template, related and not-related structures with the template were evaluated chromatographically. The highest separation factors and the imprinting factors (IF=4.73 ± 0.35 for the template) were observed in methanol, not in porogen. Only in the case of quercetin the highest IF was observed in ACN (1.88 ± 0.13). Contrary to expectations, the driving force for the affinity of the target molecules for both of polymers was enthalpic term (with an average of 54%, 82% and 84% contribution of enthalpic term for MeOH, ACN and toluene, respectively on the MIP and 53%, 57% and 65% for MeOH, ACN and toluene, respectively on the NIP). The MIP and NIP were also characterized by attenuated total reflectance analysis Fourier transform infrared spectroscopy (ATR-FTIR) and thermogravimetric analysis (TGA).


Genetic Variations in the Human Angiotensin-ConvertingEnzyme 2 and Susceptibility to Coronavirus Disease-19.
Health and economies are both affected by the coronavirus disease-19 (COVID-19) global pandemic. Angiotensin-converting enzyme 2 (ACE2) is a polymorphic enzyme that is a part of the renin-angiotensin system, and it plays a crucial role in viral entry. Previous investigations and studies revealed that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and ACE2 have a considerable association. Recently, ACE2 variants have been described in human populations in association with cardiovascular and pulmonary conditions. In this study, genetic susceptibility to COVID-19 in different populations was investigated. We evaluated the identified variants based on the predictive performance of 5 deleteriousness-scoring methods and the 2015 American College of Medical Genetics and Genomics (ACMG) guidelines. The results indicated 299 variants within the ACE2 gene. The variants were analyzed by different in-silico analysis tools to assess their functional effects. Ultimately, 5 more deleterious variants were found in the ACE2 gene. Collecting more information about the variations in binding affinity between SARS-CoV-2 and host-cell receptors due to ACE2 variants leads to progress in treatment strategies for COVID-19. The evidence accumulated in this study showed that ACE2 variants in different populations may be associated with the genetic susceptibility, symptoms, and outcome of SARS-CoV-2 infection.


Cell-type-specific and differentiation-status-dependent variations in cytotoxicity of tributyltin in cultured rat cerebral neurons and astrocytes.
Tributyltin (TBT) is an organotin used as an anti-fouling agent for fishing nets and ships and it is a widespread environmental contaminant at present. There is an increasing concern about imperceptible but serious adverse effect(s) of exposure to chemicals existing in the environment on various organs and their physiological functions, e.g. brain and mental function. Here, so as to contribute to improvement of and/or advances in in vitro cell-based assay systems for evaluating brain-targeted adverse effect of chemicals, we tried to evaluate cell-type-specific and differentiation-status-dependent variations in the cytotoxicity of TBT towards neurons and astrocytes using the four culture systems differing in the relative abundance of these two types of cells; primary neuron culture (> 95% neurons), primary neuron-astrocyte (2 : 1) mix culture, primary astrocyte culture (> 95% astrocytes), and passaged astrocyte culture (100% proliferative astrocytes). Cell viability was measured at 48 hr after exposure to TBT in serum-free medium. IC50's of TBT were 198 nM in primary neuron culture, 288 nM in primary neuron-astrocyte mix culture, 2001 nM in primary astrocyte culture, and 1989 nM in passaged astrocyte culture. Furthermore, in primary neuron-astrocyte mix culture, vulnerability of neurons cultured along with astrocytes to TBT toxicity was lower than that of neurons cultured purely in primary neuron culture. On the other hand, astrocytes in primary neuron-astrocyte mix culture were considered to be more vulnerable to TBT than those in primary or passaged astrocyte culture. The present study demonstrated variable cytotoxicity of TBT in neural cells depending on the culture condition.


Photoreduction of bacteriorhodopsin Schiff base at low humidity. A study with C13=C14 nonisomerizable artificial pigments.
The retinal protonated Schiff base of bacteriorhodopsin is photoreactive to reducing agents such as NaBH4. In the present work we have studied the effect of different protein hydration levels on the photoreductive reaction, as well as the consequences of preventing isomerization around the critical C13=C14 retinal double bond. It was revealed that the rate of light-induced NaBH4 reaction can be fitted to three phases, between 100 and 87%, from 87 to 35% and below 35% relative humidities (r.h.). The three phases are attributed to three protein regions characterized by different water affinities. Furthermore, it is shown that the PSB reduction reaction is light catalyzed even in artificial pigments derived from retinal analogs, in which isomerization around the C13=C14 double bond is prevented. It is suggested that the protein experiences light-induced conformational alterations that are not associated with C13=C14 double bond isomerization. In the 13-cis locked pigment the rate of reduction reaction is affected by r.h. levels only below 35%. The relatively low r.h. required for withdrawing water from the protein is attributed to the increased protein-water affinity in this specific pigment.


Circulating Tumour DNA Analysis for Tumour Genome Characterisation and Monitoring Disease Burden in Extramedullary Multiple Myeloma.
Mutational characterisation in extramedullary multiple myeloma (EM-MM) patients is challenging due to inaccessible EM plasmacytomas, unsafe nature of multiple biopsies and the spatial and temporal genomic heterogeneity apparent in MM (Graphical abstract). Conventional monitoring of disease burden is through serum markers and PET-CT, however these modalities are sometimes inadequate (serum markers), not performed in a timely manner (PET-CT) and uninformative for identifying mutations driving disease progression. DNA released into the blood by tumour cells (ctDNA) contains the predominant clones derived from the multiple disease foci. Blood-derived ctDNA can, therefore, provide a holistic illustration of the major drivers of disease progression. In this report, the utility of ctDNA, as an adjunct to currently available modalities in EM-MM, is presented for a patient with EM and oligosecretory (OS) disease. Whole exome sequencing of contemporaneously acquired tumour tissue and matched ctDNA samples revealed the presence of spatial and temporal genetic heterogeneity and the identification of pathways associated with drug resistance. Longitudinal monitoring of plasma samples revealed that ctDNA can be utilised to define the dynamic clonal evolution co-existent with disease progression and as an adjunct non-invasive marker of tumour burden.


When are high-tech communicators effective in Parkinson's disease?
This report describes a 63-year-old woman with Parkinson's disease showing loss of intelligibility of speech and severely impaired handwriting, despite undergoing physical and speech therapies. As the patient had sufficient residual motor abilities and adequate cognitive function and motivation, a computer-based communication aid with a software program for word prediction and voice output was tested, and was prescribed after a training period. One year later, the patient was still using the customized device to communicate and for leisure time, showing a high degree of satisfaction with the aid (assessed by QUEST 2.0), which had a positive impact on her well-being and quality of life (assessed by Psychosocial Impact of Assistive Devices Scales). In conclusion, in selected patients with Parkinson's disease, high-tech augmentative and alternative communication devices may be considered, tested, and prescribed after a positive training period. Follow-ups are necessary to monitor the effectiveness of the assistive device and respond to specific patient needs that may arise with using the device.


Calorie restriction increases muscle mitochondrial biogenesis in healthy humans.
Caloric restriction without malnutrition extends life span in a range of organisms including insects and mammals and lowers free radical production by the mitochondria. However, the mechanism responsible for this adaptation are poorly understood. The current study was undertaken to examine muscle mitochondrial bioenergetics in response to caloric restriction alone or in combination with exercise in 36 young (36.8 +/- 1.0 y), overweight (body mass index, 27.8 +/- 0.7 kg/m(2)) individuals randomized into one of three groups for a 6-mo intervention: Control, 100% of energy requirements; CR, 25% caloric restriction; and CREX, caloric restriction with exercise (CREX), 12.5% CR + 12.5% increased energy expenditure (EE). In the controls, 24-h EE was unchanged, but in CR and CREX it was significantly reduced from baseline even after adjustment for the loss of metabolic mass (CR, -135 +/- 42 kcal/d, p = 0.002 and CREX, -117 +/- 52 kcal/d, p = 0.008). Participants in the CR and CREX groups had increased expression of genes encoding proteins involved in mitochondrial function such as PPARGC1A, TFAM, eNOS, SIRT1, and PARL (all, p < 0.05). In parallel, mitochondrial DNA content increased by 35% +/- 5% in the CR group (p = 0.005) and 21% +/- 4% in the CREX group (p < 0.004), with no change in the control group (2% +/- 2%). However, the activity of key mitochondrial enzymes of the TCA (tricarboxylic acid) cycle (citrate synthase), beta-oxidation (beta-hydroxyacyl-CoA dehydrogenase), and electron transport chain (cytochrome C oxidase II) was unchanged. DNA damage was reduced from baseline in the CR (-0.56 +/- 0.11 arbitrary units, p = 0.003) and CREX (-0.45 +/- 0.12 arbitrary units, p = 0.011), but not in the controls. In primary cultures of human myotubes, a nitric oxide donor (mimicking eNOS signaling) induced mitochondrial biogenesis but failed to induce SIRT1 protein expression, suggesting that additional factors may regulate SIRT1 content during CR. The observed increase in muscle mitochondrial DNA in association with a decrease in whole body oxygen consumption and DNA damage suggests that caloric restriction improves mitochondrial function in young non-obese adults.


Parents' barriers and strategies to promote healthy eating among school-age children.
The home environment is considered one of the most important settings in regards to the development of healthy eating habits among children. The primary purpose of this study was to explore parents' barriers and strategies in promoting healthy eating in the home. The secondary objective was to determine whether the barriers and strategies parents had were different between healthy weight and overweight/obese school-age children. Semi-structured individual interviews with 14 parents of healthy weight and 11 parents of overweight/obese children (6-12 years) were conducted in family homes from August 2014 to March 2015. Transcripts were recorded and codes and themes were verified by the research team and one qualitative expert. Themes emerging from both parents of healthy weight and overweight/obese children were: 1) Parents are busy and strapped for time; 2) Cost is a barrier in providing healthy food, but parents are resourceful; 3) Children ask for junk food regularly, but parents have strategies to manage; 4) Picky eaters are a challenge but parents know they have to overcome this barrier; and 5) Early exposure to unhealthy eating influences children's food choices but strategies can help. However, parents of overweight/obese children felt a lack of support from their spouses/partners for healthy eating in the home, which was not expressed among parents of healthy weight children. Additionally, barriers and strategies were similar among parents of children from different age groups [6-9 years vs. 10-12 years (pre-adolescents)]. Our results suggest while parents faced some challenges in promoting healthy eating in the home, they utilized several strategies to overcome these barriers, which are valuable for direct intervention to improve home food environment and manage children's weight.


Cycling as a novel approach to resistance training increases muscle strength, power, and selected functional abilities in healthy older women.
Cycling on a mechanically braked cycle ergometer was used as a novel approach to compare the effects of three different 16-wk resistance-training programs on isometric force, power output, and selected functional abilities in 31 healthy 65- to 74-yr-old women. Training was conducted three times per week. During each session, individuals of the speed group performed 8 sets of 16 pedal revolutions at 40% of the maximal resistance to complete two revolutions (2 RM); strength group performed 8 sets of 8 revolutions at 80% of 2 RM; and combination group performed 4 sets of 16 revolutions at 40% and 4 sets of 8 revolutions at 80% of 2 RM. During each set, all participants were required to pedal as fast as possible with a 2-min interval between sets. All training groups significantly increased force, power, and functional abilities (maximal treadmill walking speed, vertical jumping, and box stepping) at week 8 (in the range from 6.5 to 20.8%) with no further improvement at week 16 (except maximal treadmill walking speed), but no significant differences were observed between the three groups. The novel approach to performing both low- and high-resistance training, based on the use of a cycle ergometer, has been shown to be effective in improving strength, power, and functional abilities in a group of healthy women. Even fit older women can still improve in functional abilities. Interestingly, the "high-speed" and "low-speed" programs induced an increase in both power and strength of similar magnitude.


Resolution of a Protracted Serogroup B Meningococcal Outbreak with Whole-Genome Sequencing Shows Interspecies Genetic Transfer.
A carriage study was undertaken (n = 112) to ascertain the prevalence of Neisseria spp. following the eighth case of invasive meningococcal disease in young children (5 to 46 months) and members of a large extended indigenous ethnic minority Traveller family (n = 123), typically associated with high-occupancy living conditions. Nested multilocus sequence typing (MLST) was employed for case specimen extracts. Isolates were genome sequenced and then were assembled de novo and deposited into the Bacterial Isolate Genome Sequencing Database (BIGSdb). This facilitated an expanded MLST approach utilizing large numbers of loci for isolate characterization and discrimination. A rare sequence type, ST-6697, predominated in disease specimens and isolates that were carried (n = 8/14), persisting for at least 44 months, likely driven by the high population density of houses (n = 67/112) and trailers (n = 45/112). Carriage for Neisseria meningitidis (P < 0.05) and Neisseria lactamica (P < 0.002) (2-sided Fisher's exact test) was more likely in the smaller, more densely populated trailers. Meningococcal carriage was highest in 24- to 39-year-olds (45%, n = 9/20). Evidence of horizontal gene transfer (HGT) was observed in four individuals cocolonized by Neisseria lactamica and Neisseria meningitidis One HGT event resulted in the acquisition of 26 consecutive N. lactamica alleles. This study demonstrates how housing density can drive meningococcal transmission and carriage, which likely facilitated the persistence of ST-6697 and prolonged the outbreak. Whole-genome MLST effectively distinguished between highly similar outbreak strain isolates, including those isolated from person-to-person transmission, and also highlighted how a few HGT events can distort the true phylogenetic relationship between highly similar clonal isolates.


A matter of timing: microRNA-controlled temporal identities in worms and flies.
The first microRNAs were identified in Caenorhabditis elegans based on their functions in the temporal regulation of stage-specific cell fate decisions. Until now, it was not known whether the so-called heterochronic genes that encode miRNAs are also involved in controlling developmental transitions in other organisms. New findings by Sokol et al. (this issue of Genes & Development, pp. 1591-1596) demonstrate that the Drosophila counterpart of a heterochronic miRNA gene from C. elegans, let-7, does indeed play a role in promoting stage-specific developmental events in neuromuscular tissues during the transition from larval to adult stages, thus pointing to a more widespread utilization of miRNAs in temporal regulation of animal development.


[Retention volume of some steroids during their separation by high performance liquid chromatography].
The volume of retention of corticosterone, hydrocortisone, aldosterone, dexamethasone, cortisone, methylprednisolone, and prednisolone during their separation by high-performance liquid chromatography was evaluated. The studies were performed on a Milichrome microcolumn chromatographer. The order of glucocorticoid release and volume of their retention were under study. Measurement of endogenous and exogenous plasma glucocorticoids is practically important in clinical medicine. Steroid concentrations indicate the pharmacodynamics and pharmacokinetics of exogenous glucocorticoids and their effects on the production of endogenous steroids and reflect the biorhythms of corticosterone and hydrocortisone.


[Treatment of acute nonspecific pleural empyema].
An analysis of the treatment of 240 patients with non-specific empyema of the pleura without bronchial fistula has shown the decisive significance of an adequate sanitation of the cavity by puncture or drainage. An expediency of associated use of the intrapleural antibacterial and proteolytic drugs is grounded. The general antibacterial therapy is necessary but under the threat of generalization of an infectious process and with clinical and roentgenological symptoms of the destructive process in the lungs.


Radical formation in pyrimidine bases after X, proton and alpha-particle irradiation.
Single crystals of anhydrous thymine (Ta) and cytosine monohydrate (Cm) were irradiated at room temperature using X rays, 20 MeV protons and 35 MeV alpha particles, and the relative distributions of the various radiation-induced stable radicals in the crystals were investigated. These two crystal systems were chosen because of systematic differences in their molecular packing and hydrogen-bonding network. The radicals stabilized in these systems have previously been identified and analyzed by several authors. Experimental EPR spectra could thus be reconstructed based on simulated benchmark spectra using a fitting procedure, yielding the relative amounts of the different radical species. It was found that the relative amounts of a given radical species varied with the type of radiation used, with differences being most prominent between alpha particles and protons or X rays. In Ta, an increased production of hydrogen addition or abstraction radicals was found after exposure to alpha particles. These radicals are believed to be formed predominantly from superexcited states, resulting from the higher density of ionizations along the track of the alpha particle. A corresponding reduction in radicals derived from proton transfer reactions was observed in comparison with exposure to protons or X rays. In Cm, the differences were smaller than for Ta. This is probably due to differences in the arrangement of the crystal lattice between the two systems, with Cm having an extended hydrogen-bonding network promoting fast proton transfers after ionization. Most interesting, however, was the observation that more than 50% of the radical population in Cm not could be accounted for by known radical species. The population with an unknown origin was represented with a broad singlet, having a line width of 1.5 mT and a g value of 2.0045, in the spectral reconstruction procedure.


Oat blue dwarf marafivirus resembles the tymoviruses in sequence, genome organization, and expression strategy.
The complete nucleotide sequence and genome organization of oat blue dwarf marafivirus (OBDV) were determined. The 6509 nucleotide RNA genome encodes a putative 227-kDa polyprotein (p227) with sequence motifs similar to the methyltransferase, papain-like protease, helicase, and polymerase motifs present in the nonstructural proteins of other positive strand RNA viruses. The 3' end of the open reading frame (ORF) that encodes p227 (ORF 227) also encodes the two capsid proteins: a 24-kDa capsid protein is presumably cleaved from the p227 polyprotein, whereas the 21-kDa capsid protein appears to be translated from a subgenomic RNA (sgRNA). Encoded amino acid and nucleotide sequence comparisons, as well as the OBDV genome expression strategy, show that OBDV closely resembles the tymoviruses. OBDV differs from the tymoviruses in its general biology, in its lack of a putative movement gene that overlaps the replication-associated genes, and in its fusion of the capsid gene sequences to the major ORF. OBDV also possesses a 3' poly(A) tail, as compared to the tRNA-like structures found in most tymoviral genomes. Due to the strong similarities in genome sequence and expression strategy, OBDV, and presumably the other marafiviruses, should be considered a member of the tymovirus lineage of the alpha-like plant viruses.


Molecular cloning and analysis of the ptsHI operon in Lactobacillus sake.
The ptsH and ptsI genes of Lactobacillus sake, encoding the general enzymes of the phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS), were cloned and sequenced. HPr (88 amino acids), encoded by ptsH, and enzyme I (574 amino acids), encoded by ptsI, are homologous to the corresponding known enzymes of other bacteria. Nucleotide sequence and mRNA analysis showed that the two genes are cotranscribed in a large transcript encoding both HPr and enzyme I. The transcription of ptsHI was shown to be independent of the carbon source. Four ptsI mutants were constructed by single-crossover recombination. For all mutants, growth on PTS carbohydrates was abolished. Surprisingly, the growth rates of mutants on ribose and arabinose, two carbohydrates which are not transported by the PTS, were accelerated. This unexpected phenotype suggests that the PTS negatively controls ribose and arabinose utilization in L. sake by a mechanism different from the regulation involving HPr described for other gram-positive bacteria.


Foreign signal peptides can constitute a barrier to functional expression of periplasmic proteins in Haemophilus influenzae.
To study the periplasmic branch of iron (ferric ion) uptake systems in Gram-negative bacteria, genetic reconstitution experiments were initiated in Haemophilus influenzae involving exchange of the periplasmic iron-binding protein. The expression of many of the heterologous periplasmic ferric-binding proteins (FbpAs) was quite limited. Transformation experiments with the fbpA gene from Neisseria gonorrhoeae yielded two colony sizes with different phenotypic characteristics. The small colonies contained the intact N. gonorrhoeae fbpA gene and were deficient in utilization of transferrin iron. The large colonies contained hybrid H. influenzae/N. gonorrhoeae fbpA genes, were proficient in transferrin iron utilization and had enhanced levels of expression of FbpA. These hybrid genes included several that encoded the mature N. gonorrhoeae FbpA with the H. influenzae signal peptide. To more fully evaluate the effect of foreign signal peptides, a series of hybrid genes were prepared that exchanged the signal peptides from H. influenzae FbpA, N. gonorrhoeae FbpA and the TEM-1 beta-lactamase. The presence of the H. influenzae leader was required for functional expression of FbpAs and was shown to dramatically increase the level of beta-lactamase activity.


H-Scan Discrimination for Tumor Microenvironmental Heterogeneity in Melanoma.
Melanoma is a form of malignant skin cancer that exhibits significant inter-tumoral differences in the tumor microenvironment (TME) secondary to genetic mutations. The heterogeneity may be subtle but can complicate the treatment of metastatic melanoma, contributing to a high mortality rate. Therefore, developing an accurate and non-invasive procedure to discriminate microenvironmental heterogeneity to facilitate therapy selection is an important goal. In vivo murine melanoma models that recapitulate human disease using synchronous implanted YUMM 1.7 (Yale University Mouse Melanoma) and YUMMER 1.7 (Yale University Mouse Melanoma Exposed to Radiation) murine melanoma lines were investigated. Mice were treated with antibodies to modulate the immune response and longitudinally scanned with ultrasound (US). US radiofrequency data were processed using the H-scan analysis, attenuation estimation and B-mode processing to extract five US features. The measures were used to compare different TMEs (YUMMER vs. YUMM) and responses to immunomodulatory therapies with CD8 depletion or programmed cell death protein 1 (PD-1) inhibition. Multiparametric analysis produced a combined H-scan parameter, resolving significant differences (i) between untreated YUMMER and YUMM and (ii) between untreated, PD-1-treated and CD8-treated YUMMER. However, more importantly, the B-mode and attenuation measures failed to differentiate YUMMER and YUMM and to monitor treatment responses, indicating that H-scan is required to differentiate subtle differences within the TME. We anticipate that the H-scan analysis could discriminate heterogeneous melanoma metastases and guide diagnosis and treatment selection, potentially reducing the need for invasive biopsies or immunologic procedures.


Fast, anticipatory smooth-pursuit eye movements appear to depend on a short-term store.
Anticipatory smooth pursuit before the expected appearance of a moving target can reduce the initial retinal blur caused by the 100-ms delay of visual feedback. Humans, though, can only voluntarily generate smooth velocities up to about 5 degrees/s without a moving target. However, previous experiments have shown that repetitive brief presentations of a moving target every few seconds appear to charge an internal store, the contents of which can later be released to generate higher velocity anticipatory movements. This store's longevity was assessed here by repetitively presenting a moving target for 500 ms at different known intervals up to 7.2 s. Target motion at 25 degrees/s or 50 degrees/s was tested, with presentations in alternate directions or the same direction. Anticipatory velocity, measured 100 ms after target onset, decreased with increasing interval for all target motion conditions. A decrease was still seen when accurate timing cues were given before each presentation, suggesting that the drive for anticipatory pursuit is held in a short-term store lasting a few seconds which can enhance the low velocities produced by volition alone. The results also demonstrate that high-velocity anticipatory pursuit helps to overcome the temporal delays in the system and allows target velocity to be matched at an earlier time.


Short-day-like body weight changes do not prevent fat pad compensation after lipectomy in Siberian hamsters.
Long-day (LD)-housed Siberian hamsters show compensatory increases in white adipose tissue (WAT) weight after lipectomy, whereas hamsters exposed to short days (SDs) for a long duration (22 wk) do not. We tested whether SD-induced body weight changes prevent fat pad compensation after lipectomy. In experiment 1, hamsters with lesions of the paraventricular nucleus of the hypothalamus (PVNx) rapidly increased body weight similarly to 22-wk SD-exposed hamsters. In experiment 2, LD-housed hamsters were food restricted for 22 wk and then pair fed with SD-housed hamsters for 12 wk to produce body weight changes mimicking those of ad libitum-fed SD-exposed animals. Epididymal WAT (EWAT) lipectomy (EWATx) of PVNx or food-restricted hamsters elicited compensatory increases in retroperitoneal and inguinal WAT (RWAT and IWAT) weights. Unlike other fat pads, EWAT was less affected by food restriction or PVNx than by SD exposure. In general, food restriction decreased adipocyte number, whereas SD exposure decreased adipocyte size. PVNx increased RWAT adipocyte size and IWAT adipocyte number. These results suggest that the lack of body fat compensation by EWATx hamsters exposed to SDs for a long duration is due to SD-associated responses other than body weight changes per se.


Modeling and analysis of individualized pharmacokinetics and pharmacodynamics for volatile anesthesia.
The presented procedure aims to establish an in-depth understanding of a derived mathematical model for drug distribution, pharmacokinetics, and drug effect, pharmacodynamics, during volatile anesthesia. A physiologically based, patient-specific model is derived, where the pharmacokinetic (PK) part consists of multiple blood and tissue compartmental models, each adjusted to the weight, height, gender, and age of the patient. The pharmacodynamic (PD) part is described by an effect site compartment and the Hill equation both linking the hypnotic effect measured by the Bispectral Index (BIS) to the arterial anesthetic concentration. Via a global sensitivity analysis the patient-specific PK and PD variables and parameters are analyzed regarding their influence on the measurable outputs, which are the end-tidal concentration of the volatile anesthetic and the BIS. Via this analysis, the uncertainty introduced by PD variability is identified to be more significant than the uncertainty introduced by PK variability. A case study of isoflurane-based anesthesia shows that the simulation results of the individualized PK variables are in good accordance with the measured end-tidal concentration. However, the PD parameters need to be estimated online to predict the hypnotic depth, measured by the BIS, correctly. As a result of this study, the aim should be to focus on the individual identification of the PD parameters before and during anesthesia with future application in safe and robust model predictive control.


The effect of behavioral stimulant doses of amphetamine on blood pressure.
Blood pressure responses to amphetamine have been studied both in humans and in rats. Blood pressure was monitored in patients who had experienced an adverse behavioral reaction to the drug. All of the patients had self-administereed amphetamine, and later sought medical attention for their adverse reaction. In a series of 14 patients, there was no evidence that amphetamine had evoked a sustained increase in blood pressure. In rats, experiments were conducted in two steps: (1) a determination of the doses of amphetamine that cause behavioral stimulation, and (2) an evaluation of the blood pressure effects of the same dose of amphetamine. In control animals, behavioral stimulant doses of amphetamine exerted only transient effects on blood pressure. In pithed animals, ie, animals devoid of all central mechanisms, amphetamine exerted a sustained effect on blood pressure. It is concluded that the potential ability of amphetamine to evoke sustained cardiovascular responses is damped by the central nervous system of intact animals.


Investigation on the inactivation effect and mechanism of Listeria monocytogenes in fresh-cut cucumber during storage by ultrasound combined with sodium hypochlorite.
Fresh agricultural products are frequently contaminated with Listeria monocytogenes (L. monocytogenes), which threatens consumer health. The mechanism of the inhibitory effect of ultrasound and sodium hypochlorite (US-NaClO) on L. monocytogenes on fresh-cut cucumber remains poorly understood. Therefore, the bactericidal ability and mechanism of US-NaClO treatment on L. monocytogenes were studied on fresh-cut cucumber during storage using various approaches such as determination of intracellular material leakage, scanning electron microscopy, flow cytometry, and expression analysis of virulence genes. The results showed that the number of L. monocytogenes on fresh-cut cucumber was significantly reduced after ultrasound treatment for 5 min in combined with 75 ppm sodium hypochlorite treatment(P < 0.05). The US-NaClO treatment affected cell morphology, impaired cell membrane integrity, increased cell membrane permeability, and reduced the concentration of K+, inorganic phosphate, ATP, proteins, and DNA in bacterial cells, leading to the inactivation of microorganisms. In addition, the US-NaClO treatment downregulated expression of the virulence genes actA, hly, inlA, mpl, pclA, and plcB, thus decreasing the pathogenicity of bacteria. It can avoid contamination by pathogenic bacteria during the production of fresh-cut cucumber, while providing safety assurance for production.


Age of onset of substance use in patients with dual diagnosis and its association with clinical characteristics, risk behaviors, course, and outcome: A retrospective study.
Do patients with dual diagnosis (DD) with an early initiation of substance use and subsequent early onset substance use disorder (EOS) differ from those with late onset substance use disorder (LOS) regarding characteristics, sex-related risk behaviors, course, and outcome? The aim is to study EOS and LOS groups of patients with DD attending a treatment center with regard to clinical characteristics, type of psychiatric disorders, risk behaviors, and short-term outcome. Retrospective chart review of patients diagnosed with DD (n = 307) with regard to the above variables. Diagnoses of both substance use disorders (SUDs) and psychiatric disorders were made by qualified psychiatrists as per the International Classification of Diseases, 10th revision. Among 307 participants with DD, 100 were in EOS group (onset of SUD before 18 years of age as assessed clinically) and 207 in LOS group. Cannabis as the primary substance was more prevalent in the EOS (30%) than that of the LOS group (12%). Psychotic disorders were more prevalent in the EOS group (41%) followed by mood disorders (30%), while the reverse was true in the LOS group (27.5% and 56.5%, respectively). When compared to the LOS group, the EOS group had higher number of psychiatric admissions, reported higher prevalence of risky sexual behaviors (unprotected sexual intercourse, multiple sexual partners, and history of sexual intercourse with commercial sex workers), showed poorer treatment adherence, and worse outcome regarding both psychiatric disorder and SUD (all differences significant at P < 0.05). DD patients differ significantly based on the age of onset of substance use. These may have therapeutic and management implications.


Detection of lymph nodes micrometastases in Dukes' A and B colorectal cancer using anti-cytokeratin antibodies AE1/AE3.
To detect lymph nodes micrometastases and analyze its correlation with clinicopathological parameters in Dukes' A and B colorectal cancer patients. One hundred and fourteen patients with colorectal cancer (Dukes' A 16; Dukes' B 98) undergoing curative operation without histological lymph nodes metastases were studied between 2001 and 2003. A total of 2,481 lymph nodes were analyzed using monoclonal cytokeratin antibody AE1/AE3 (DAKO, Carpinteria, CA) for immunohistochemistry. In total, 33 (29%) patients were positive for cancer cell by immunohistochemistry. In 31 (94%) patients of them positive nodes showed single tumor cell or small groups of tumor cells; and tumor deposits measuring 0.2 and 0.37 mm in diameter in another 2 (6%) patients. Micrometastases were mainly located in the subcapsular sinus or paracortical sinus. There was no correlation between the positive lymph nodes and gender, age, tumor site, tumor size, histological type, histological grade, invasion depth, Dukes' staging and microsatellite instability (P>0.05). Our findings suggest that immunohistochemical technique using monoclonal cytokeratin antibody AE1/AE3 may be a sensitive and reliable method for detecting lymph nodes micrometastases in Dukes' A and B colorectal cancer. The clinical significance of lymph nodes micrometastases is still not confirmed.


Crenobacter cavernae sp. nov., isolated from a karst cave, and emended description of the genus Crenobacter.
A Gram-stain-negative, rod-shaped, motile and strictly aerobic novel bacterial isolate, designated strain K1W11S-77T, was obtained from a water sample that was collected from a karst cave in Guizhou province, PR China. The results of a phylogenetic analysis based on 16S rRNA gene sequences indicated that K1W11S-77T represented a member of the genus Crenobacter within the family Neisseriaceae of the phylum Proteobacteria. K1W11S-77T was phylogenetically closely related to Crenobacter luteusYIM 78141T (Their 16S rRNA gene sequence similarity is 95.02 %). Growth of K1W11S-77T occurred at 10-30 °C, at pH 7-9, and in the presence of 0-1 % (w/v) NaCl. The major cellular fatty acids were C12 : 0, C16 : 0, C18:1ω7c and summed feature 3. The major isoprenoid quinone was Q-8. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and one unidentified phospholipid. The genome of K1W11S-77T was 3.27 Mb long and encoded 3167 annotated genes. The DNA G+C content of the genomic DNA was 65.3 mol%. On the basis of phylogenetic, phenotypic and chemotaxonomic characteristics, K1W11S-77T is considered to represent a novel species of the genus Crenobacter, for which the name Crenobactercavernae sp. nov. is proposed. The type strain is K1W11S-77T (=CGMCC 1.13527T=NBRC 113452T).


Influence of dietary protein and carbohydrate sources on nitrogen metabolism and carbohydrate fermentation by ruminal microbes in continuous culture.
Four diets containing 15% CP were formulated to study the effects of dietary carbohydrate and protein sources on N metabolism and carbohydrate fermentation by ruminal bacteria. Diets were supplied to eight dual-flow continuous culture fermenters during three experimental periods in a randomized complete block design. Six replications were obtained for each diet. Treatments were arranged as a 2 X 2 factorial with two carbohydrate and two protein sources. Carbohydrate sources were corn and barley and protein sources were soybean meal (SBM) and fish meal (FM). Approximately 40% of the dietary CP was derived from SBM or FM and corn or barley provided 39% of dietary DM. All diets contained 15% grass hay, 20% wheat straw, and 10.1 to 15.3% solka floc (DM basis). Interactions (P less than .05) were observed between dietary carbohydrate and protein sources, resulting in a depression of VFA production (moles/day) and digestion (percentage) of ADF and cellulose when the corn-FM diet was fed. True OM digestion (percentage) was higher (P less than .05) for SBM than for FM diets and for corn than for barley diets. Although dietary CP degradation (percentage) was higher (P less than .05) for SBM than for FM diets, non-NH3 N in the effluent (grams/day) was not different among diets due to a greater (P less than .05) bacterial N flow for SBM than for FM diets. Despite the lower amino acid (AA) intake (P less than .05) for corn than for barley diets and also for FM than for SBM diets, flows (grams/day) of total AA, essential AA (EAA), and nonessential AA (NEAA) were similar (P greater than .05) among diets. However, greater (P less than .05) total AA, EAA, and NEAA flows (percentage of AA intake) were found for corn than for barley diets and for FM than for SBM diets. It is concluded, therefore, that ruminal escape protein derived from corn or FM has a significant effect on manipulating AA leaving the ruminal fermentation.


Shoal sex composition and predation risk influence sub-adult threespine stickleback shoaling decisions.
Group living reduces individual predation risk most effectively when group members are behaviorally and phenotypically similar. Group preferences are influenced by the individual, the members of the shoal, and the environmental conditions. While shoaling behavior has been studied extensively in the threespine stickleback (Gasterosteus aculeatus), it is unclear whether the sex of shoal mates influences the shoal preference of non-reproductive males and females and how this changes under increasing predation risk. Although non-reproductively active sticklebacks are sexually monochromatic in appearance, sex-related differences may result in sexual segregation when shoaling. Here we show that male and female sub-adult threespine sticklebacks had contrasting preferences for shoal mate sex, and that this preference was dependent on the level of predation risk during standardized experimental choice tests. In detail, test fish shoal with the opposite sex within low predation risk trials and with same-sex shoals within high predation risk trials. This difference might be linked to activity patterns; test males were more active than females. Our results demonstrate that differences between the sexes in a species with a sexually monochromatic non-reproductive stage can result in sex-related shoaling preferences. Most studies examining sexual segregation focus on sexually dimorphic species, but these results highlight the potentially widespread occurrence of sexual segregation beyond the sexually dimorphic reproductive stage.


[Unusual causes of gait disorders].
Gait disorder is a common complaint in general practice, in particular in the elderly, and more than one cause frequently interact to produce various abnormalities of gait, including neurological, orthopedic, rhumatologic or ophthalmologic conditions, among others. The most frequent etiologies of gait impairment include sensory deficits, cervical myelopathy, vascular encephalopathy, parkinsonism and normal pressure hydrocephalus. In a particular individual, several of them may contribute to alter gait and, as a general rule, the treatment of these well-established conditions is generally considered difficult. Besides them, a number of rare and underrecognized neurological conditions may also produce a gait disorder as their initial, main or exclusive feature, including dopa-responsive dystonia, stiff-person syndrome, orthostatic tremor, primary progressive freezing gait and cursive epilepsy. In this review article, the clinical, therapeutic and other characteristics of these conditions are revisited and a typical illustrative case is reported for each. Since most of these conditions are responsive to specific treatments, we believe that a better knowledge of these unusual forms of gait disorder may allow general practitioners to identify them more accurately, to assess them and to improve therapeutic strategies.


Profiling of repetitive RNA sequences in the blood plasma of patients with cancer.
Liquid biopsies provide a means for the profiling of cell-free RNAs secreted by cells throughout the body. Although well-annotated coding and non-coding transcripts in blood are readily detectable and can serve as biomarkers of disease, the overall diagnostic utility of the cell-free transcriptome remains unclear. Here we show that RNAs derived from transposable elements and other repeat elements are enriched in the cell-free transcriptome of patients with cancer, and that they serve as signatures for the accurate classification of the disease. We used repeat-element-aware liquid-biopsy technology and single-molecule nanopore sequencing to profile the cell-free transcriptome in plasma from patients with cancer and to examine millions of genomic features comprising all annotated genes and repeat elements throughout the genome. By aggregating individual repeat elements to the subfamily level, we found that samples with pancreatic cancer are enriched with specific Alu subfamilies, whereas other cancers have their own characteristic cell-free RNA profile. Our findings show that repetitive RNA sequences are abundant in blood and can be used as disease-specific diagnostic biomarkers.


Regulation of Hippo signaling by EGFR-MAPK signaling through Ajuba family proteins.
EGFR and Hippo signaling pathways both control growth and, when dysregulated, contribute to tumorigenesis. We find that EGFR activates the Hippo pathway transcription factor Yorkie and demonstrate that Yorkie is required for the influence of EGFR on cell proliferation in Drosophila. EGFR regulates Yorkie through the influence of its Ras-MAPK branch on the Ajuba LIM protein Jub. Jub is epistatic to EGFR and Ras for Yorkie regulation, Jub is subject to MAPK-dependent phosphorylation, and EGFR-Ras-MAPK signaling enhances Jub binding to the Yorkie kinase Warts and the adaptor protein Salvador. An EGFR-Hippo pathway link is conserved in mammals, as activation of EGFR or RAS activates the Yorkie homolog YAP, and EGFR-RAS-MAPK signaling promotes phosphorylation of the Ajuba family protein WTIP and also enhances WTIP binding to the Warts and Salvador homologs LATS and WW45. Our observations implicate the Hippo pathway in EGFR-mediated tumorigenesis and identify a molecular link between these pathways.


Fasting decreases thyrotropin responsiveness to thyrotropin-releasing hormone: a potential cause of misinterpretation of thyroid function tests in the critically ill.
We have previously reported that caloric deprivation inhibits peripheral T4 metabolism and blunts the TSH response to TRH in euthyroid obese subjects. To determine whether these phenomena also occur in hypothyroid subjects, T4, T3, rT3, and the TSH response to TRH were measured initially and after a 60-h fast in seven hypothyroid patients. Short term fasting caused a 29% decrement in the maximum serum TSH increment and a 32% decrement in the integrated TSH response to TRH (P less than 0.01). In two subjects with mild hypothyroidism, basal TSH as well as the TSH response to TRH were reduced to levels within the normal range. Specifically, basal TSH values decreased from 7.6 to 3.5 microU/ml and from 11 to 4.1 microU/ml. In the seven subjects, mean serum T3 decreased significantly from 88 to 60 ng/dl, (P less than 0.05) and rT3, initially undetectable in six of seven subjects, rose to detectable or low normal values in four of seven subjects, serum T4 remained at 2.7 micrograms/dl during both study periods. We conclude that 1) fasting induces changes in both peripheral thyroid hormone metabolism and the hypothalamic-pituitary axis in hypothyroid individuals which are qualitatively similar to those that occur in euthyroid subjects; and 2) in certain hypothyroid subjects, fasting alone can decrease basal TSH values to within the normal range. If these data can be extrapolated to critically ill subjects whose caloric intake may be diminished, they suggest that basal TSH concentrations in moderately and severely hypothyroid critically ill subjects will accurately reflect the biochemically hypothyroid state. However, mild degrees of hypothyroidism in critically ill subjects might be overlooked due to the lowering effect of fasting or poor caloric intake alone on basal TSH concentrations.


Negative soil feedbacks accumulate over time for non-native plant species.
The enemy release hypothesis is a common explanation for species invasions, suggesting that introduced species benefit from leaving behind natural enemies in the native range. However, any such advantage may attenuate over time. In this study, we test a prediction of this more dynamic enemy release hypothesis: that non-native plant species that became established longer ago exhibit stronger negative feedbacks with the soil. Consistent with declining enemy release over time, we found increasingly negative soil feedbacks for species established longer ago in New Zealand. Negative soil feedbacks were also stronger for more widespread species, but weaker for more locally abundant species, suggesting that species accumulate negative interactions as they spread and can be locally regulated by these interactions. We also present data to support the common assumption that relatives have similar impacts on and responses to soil communities. Together, these data highlight the dynamic nature of novel interactions arising from species introductions.


Obstetric complications with GDM. Effects of maternal weight.
Obstetric complications recorded prospectively were assessed retrospectively in 150 women with gestational diabetes mellitus (GDM) and 305 control subjects matched for age, parity, and ethnicity. Intensive diet therapy and self-monitoring of capillary blood glucose were used to obtain postprandial euglycemia; 22% of GDM subjects required insulin. GDM and control subjects were grouped by body mass index to detect any influence of maternal prepregnancy weight on outcome. Polyhydramnios, preterm labor, and pyelonephritis were not more frequent in GDM, but hypertension without proteinuria (7.3 vs. 3.3%) and preeclampsia (8 vs. 3.9%) were more frequent in GDM. The frequency of hypertensive complications in GDM was not totally attributable to being overweight. Abnormalities of labor, birth trauma, and fetal macrosomia were not more common in GDM; 6.7% of the infants of mothers with GDM weighed greater than 4200 g at birth compared with 3.6% of control infants (NS), and 10% were large for gestational age and sex compared with 6.6% of control infants (NS). Despite this, cesarean delivery was more common in GDM (35.3 vs. 22%, P less than 0.01), mostly due to significantly more cesarean births without labor.


A new hypothesis for foregut and heart tube formation based on differential growth and actomyosin contraction.
For decades, it was commonly thought that the bilateral heart fields in the early embryo fold directly towards the midline, where they meet and fuse to create the primitive heart tube. Recent studies have challenged this view, however, suggesting that the heart fields fold diagonally. As early foregut and heart tube morphogenesis are intimately related, this finding also raises questions concerning the traditional view of foregut formation. Here, we combine experiments on chick embryos with computational modeling to explore a new hypothesis for the physical mechanisms of heart tube and foregut formation. According to our hypothesis, differential anisotropic growth between mesoderm and endoderm drives diagonal folding. Then, active contraction along the anterior intestinal portal generates tension to elongate the foregut and heart tube. We test this hypothesis using biochemical perturbations of cell proliferation and contractility, as well as computational modeling based on nonlinear elasticity theory including growth and contraction. The present results generally support the view that differential growth and actomyosin contraction drive formation of the foregut and heart tube in the early chick embryo.


Magnolol Inhibits the Growth of Non-Small Cell Lung Cancer via Inhibiting Microtubule Polymerization.
The tubulin/microtubule system, which is an integral component of the cytoskeleton, plays an essential role in mitosis. Targeting mitotic progression by disturbing microtubule dynamics is a rational strategy for cancer treatment. Microtubule polymerization assay was performed to examine the effect of Magnolol (a novel natural phenolic compound isolated from Magnolia obovata) on cellular microtubule polymerization in human non-small cell lung cancer (NSCLC) cells. Cell cycle analysis, mitotic index assay, cell proliferation assay, colony formation assay, western blotting analysis of cell cycle regulators, Annexin V-FITC/PI staining, and live/dead viability staining were carried out to investigate the Magnolol's inhibitory effect on proliferation and viability of NSCLS cells in vitro. Xenograft model of human A549 NSCLC tumor was used to determine the Magnolol's efficacy in vivo. Magnolol treatment effectively inhibited cell proliferation and colony formation of NSCLC cells. Further study proved that Magnolol induced the mitotic phase arrest and inhibited G2/M progression in a dose-dependent manner, which were mechanistically associated with expression alteration of a series of cell cycle regulators. Furthermore, Magnolol treatment disrupted the cellular microtubule organization via inhibiting the polymerization of microtubule. We also found treatment with NSCLC cells with Magnolol resulted in apoptosis activation through a p53-independent pathway, and autophgy induction via down-regulation of the Akt/mTOR pathway. Finally, Magnolol treatment significantly suppressed the NSCLC tumor growth in mouse xenograft model in vivo. These findings identify Magnolol as a promising candidate with anti-microtubule polymerization activity for NSCLC treatment.


Interior micro-CT with an offset detector.
The size of field-of-view (FOV) of a microcomputed tomography (CT) system can be increased by offsetting the detector. The increased FOV is beneficial in many applications. All prior investigations, however, have been focused to the case in which the increased FOV after offset-detector acquisition can cover the transaxial extent of an object fully. Here, the authors studied a new problem where the FOV of a micro-CT system, although increased after offset-detector acquisition, still covers an interior region-of-interest (ROI) within the object. An interior-ROI-oriented micro-CT scan with an offset detector poses a difficult reconstruction problem, which is caused by both detector offset and projection truncation. Using the projection completion techniques, the authors first extended three previous reconstruction methods from offset-detector micro-CT to offset-detector interior micro-CT. The authors then proposed a novel method which combines two of the extended methods using a frequency split technique. The authors tested the four methods with phantom simulations at 9.4%, 18.8%, 28.2%, and 37.6% detector offset. The authors also applied these methods to physical phantom datasets acquired at the same amounts of detector offset from a customized micro-CT system. When the detector offset was small, all reconstruction methods showed good image quality. At large detector offset, the three extended methods gave either visible shading artifacts or high deviation of pixel value, while the authors' proposed method demonstrated no visible artifacts and minimal deviation of pixel value in both the numerical simulations and physical experiments. For an interior micro-CT with an offset detector, the three extended reconstruction methods can perform well at a small detector offset but show strong artifacts at a large detector offset. When the detector offset is large, the authors' proposed reconstruction method can outperform the three extended reconstruction methods by suppressing artifacts and maintaining pixel values.


Trends in total knee replacement surgeries and implications for public health, 1990-2000.
Total joint replacements are important surgical interventions for treating severe arthritis of weight-bearing joints. The most common indication for total knee replacement (TKR) is osteoarthritis of the knee joint. The goals of this study were to assess the trend in rate of TKR in Wisconsin and to describe the economic impact of these surgical procedures on the health care system. A population-based cross-sectional study of TKR surgeries was conducted among Wisconsin residents aged > or = 45 years. The Wisconsin inpatient hospital discharge data from 1990 through 2000 were used. Rates were age-adjusted to the 2000 U.S. population, and charges for TKR were adjusted for inflation. From 1990 through 2000, the age-adjusted rate for TKR increased by 81.5% (from 162 to 294 per 100,000; p<0.001). The rate increased the most among the youngest age group (45-49 years), rate ratio 5.1 for men, 4.2 for women. The total charges for TKR increased from 69.4 million dollars to 148 million dollars (109.2% inflation-adjusted increase). Medicare received the highest proportion of charges for TKR procedures, but throughout the study period, the proportion of charges covered by private insurance increased by 39%. The rate and costs of TKR procedures among Wisconsin residents increased substantially from 1990 through 2000, especially among younger age groups. Changes in medical practices probably accounted for some of this increase, but these trends also may reflect an increased prevalence of osteoarthritis, which in turn may be related to dramatic increases in the number of individuals who are overweight.


[Dilatation of benign colo-colonic and colo-rectal anastomotic stenosis with radiology-guided balloon catheter].
Eleven patients with benign postoperative colonic strictures were treated with balloon dilatation November, 1990, through November, 1995. The anastomosis was sigmoidrectal in 7 patients and colocolic in 4 patients. All patients were submitted to contrast enema and colonscopy to assess the site, shape, grade and length of the stenosis. Biopsy was performed in 7 patients whose strictures had developed 2 months or more postoperatively. The dilatation was performed with 20-mm balloon catheters in the strictures developed 30 days postoperatively, to avoid any complications, and with 30-mm balloon catheters in the other cases. Balloon dilatation was performed under fluoroscopic guidance, with no-drug treatment. The procedure was well tolerated by all patients. One or two dilatation sessions were performed in ten and one patients, respectively. No complications were observed. The results were satisfactory in all cases, with symptom resolution. Follow-up included clinical, endoscopic and radiologic assessment. At follow-up, the technical result was good in all patients and the symptoms were completely relieved. In our experience, radiologically-guided balloon catheter dilatation proved to be an easy, safe and effective tool to treat benign postoperative colonic strictures.


Kinetics and physiologic relevance of the inactivation of alpha 1-proteinase inhibitor, alpha 1-antichymotrypsin, and antithrombin III by matrix metalloproteinases-1 (tissue collagenase), -2 (72-kDa gelatinase/type IV collagenase), and -3 (stromelysin).
Serpins encompass a superfamily of proteinase inhibitors that regulate many of the serine proteinases involved in inflammation and hemostasis. In vitro, many serpins are catalytically inactivated by proteinases that they do not inhibit, leading to the concept of proteolytic down-regulation of serpin inhibitory capacity. The extent to which down-regulation of serpin activity occurs in vivo is debated, since little is known of the rates at which the process occurs. To address this debate, we have measured the rates of inactivation of three serpins, alpha 1-proteinase inhibitor (alpha 1PI), alpha 1-antichymotrypsin (alpha 1ACT), and antithrombin III (ATIII), by three human matrix metalloproteinases (MMPs-1, -2, and -3) thought to be involved in tissue destruction and repair. Our object was to establish a working kinetic model which can be used to predict whether serpin inactivation by these proteinases is likely to occur in vivo. We determined the rates of inactivation of these three serpins by each of the MMPs and compared these to rates of inhibition of the MMPs by an endogenous inhibitor, alpha 2-macroglobulin. An equation designed to predict the extent of substrate hydrolyzed by an enzyme in the presence of an enzyme inhibitor gave the following predictions of the inactivation in vivo: (i) ATIII is unlikely to be inactivated by the MMPs. (ii) MMP-2 (72-kDa gelatinase/type IV collagenase) is unlikely to inactivate any of the three serpins. (iii) MMP-1 (tissue collagenase) will inactivate alpha 1PI and alpha 1ACT only when its concentration saturates that of its controlling inhibitors. (iv) MMP-3 (stromelysin) may inactivate small amounts of alpha 1PI and more significant amounts of alpha 1ACT, even in the presence of its controlling inhibitors. Any physiologic or pathologic inactivation of these serpins by these MMPs that occurs in vivo will probably be due to MMP-3, and will likely only take place in tissues and inflammatory loci where the concentration of MMP inhibitors is depressed.


Guidance on conducting systematic reviews/meta-analyses of pharmacoepidemiological studies of safety outcomes: the gap is now filled.
Until recently, no comprehensive guidance specifically on the conduction of systematic reviews and meta-analyses of pharmacoepidemiological studies of safety outcomes was available. In December 2015, the European Network of Centres for Pharmacoepidemiology and Pharamacovigilance (ENCePP), a network coordinated by the European Medicines Agency, published their 'Guidance on conducting systematic reviews and meta-analyses of completed comparative pharmacoepidemiological studies of safety outcomes', filling an important gap in the field. This paper highlights the ENCePP recommendations in terms of study identification, data extraction, study quality appraisal and analytical plan. Although the ENCePP document should not be considered as definitive, since it will likely be refined following researchers' feedback, it is expected that it will be highly influential and useful for the field, with the ultimate goal to improve and standardise the conduction and reporting of systematic reviews/meta-analyses of pharmacoepidemiological studies of safety outcomes.


Assessing Ploidy Level Analysis and Single Pollen Genotyping of Diploid and Euploid Citrus Genotypes by Fluorescence-Activated Cell Sorting and Whole-Genome Amplification.
Flow cytometry is widely used to determine genome size and ploidy level in plants. This technique, when coupled with fluorescence-activated cell sorting (FACS), whole genome amplification and genotyping (WGA), opens up new opportunities for genetic studies of individualized nuclei. This strategy was used to analyze the genetic composition of single pollen nuclei of different citrus species. The flow cytometry and microscope observations allowed us to differentiate the populations of pollen nuclei present in the diploid and euploid genotypes analyzed, showing that citrus has binuclear pollen. We have identified in the "CSO" tangor an additional nuclei population composed by the vegetative plus generative nuclei. Genotyping of this nuclei population revealed that vegetative and generative nuclei show the same genetic configuration. In addition, we have demonstrated the presence of unreduced gametes in the diploid genotype "Mexican lime." Genomic amplification is a robust method for haploid nuclei genotyping with several molecular markers, whereas in diploid nuclei using heterozygous markers showed a bias towards one of the two alleles, limiting the use of this tool in this type of nuclei. We further discuss the importance and applications of single pollen genotyping in citrus genetic studies.


Lidocaine Prevents Oxidative Stress-Induced Endothelial Dysfunction of the Systemic Artery in Rats With Intermittent Periodontal Inflammation.
Periodontal inflammation causes endothelial dysfunction of the systemic artery. However, it is unknown whether the use of local anesthetics during painful dental procedures alleviates periodontal inflammation and systemic endothelial function. This study was designed to examine whether the gingival or systemic injection of lidocaine prevents oxidative stress-induced endothelial dysfunction of the systemic artery in rats with intermittent periodontal inflammation caused by lipopolysaccharides (LPS). Some rats received 1500 µg LPS injections to the gingiva during a week interval from the age of 8 to 11 weeks (LPS group). Lidocaine (3 mg/kg), LPS + lidocaine (3 mg/kg), LPS + lidocaine (1.5 mg/kg), and LPS + lidocaine (3 mg/kg, IP) groups simultaneously received gingival 1.5 or 3 mg/kg or IP 3 mg/kg injection of lidocaine on the same schedule as the gingival LPS. Isolated aortas or mandibles were subjected to the evaluation of histopathologic change, isometric force recording, reactive oxygen species, and Western immunoblotting. Mean blood pressure and heart rate did not differ among the control, LPS, LPS + lidocaine (3 mg/kg), and lidocaine (3 mg/kg) groups. LPS application reduced acetylcholine (ACh, 10 to 10 mol/L)-induced relaxation (29% difference at ACh 3 × 10 mol/L, P = .01), which was restored by catalase. Gingival lidocaine (1.5 and 3 mg/kg) dose dependently prevented the endothelial dysfunction caused by LPS application (24.5%-31.1% difference at ACh 3 × 10 mol/L, P = .006 or .001, respectively). Similar to the gingival application, the IP injection of lidocaine (3 mg/kg) restored the ACh-induced dilation of isolated aortas from rats with the LPS application (27.5% difference at ACh 3 × 10 mol/L, P < .001). Levels of reactive oxygen species were double in aortas from the LPS group (P < .001), whereas the increment was abolished by polyethylene glycol-catalase, gingival lidocaine (3 mg/kg), or the combination. The LPS induced a 4-fold increase in the protein expression of tumor necrosis factor-α in the periodontal tissue (P < .001), whereas the lidocaine (3 mg/kg) coadministration partly reduced the levels. Lidocaine application also decreased the protein expression of the nicotinamide adenine dinucleotide phosphate oxidase subunit p47phox, which was enhanced by the gingival LPS (5.6-fold increase; P < .001). Lidocaine preserved the aortic endothelial function through a decrease in arterial reactive oxygen species produced by nicotinamide adenine dinucleotide phosphate oxidase and periodontal tumor necrosis factor-α levels in rats with periodontal inflammation. These results suggest the beneficial effect of the gingival application of local anesthetics on the treatment of periodontal diseases on endothelial function of systemic arteries.