Hippocampal vulnerability and subacute response following varied blast magnitudes.
Clinical outcomes from blast neurotrauma are associated with higher order cognitive functions such as memory, problem solving skills and attention. Current literature is limited to a single overpressure exposure or repeated exposures at the same level of overpressure and is focused on the acute response (<3 days). In an attempt to expand the understanding of neuropathological and molecular changes of the subacute response (7 days post injury), we used an established rodent model of blast neurotrauma. Three pressure magnitudes (low, moderate and high) were used to evaluate molecular injury thresholds. Immunohistochemical analysis demonstrated increased cleaved caspase-3 levels and loss of neuronal population (NeuN+) within the hippocampus of all pressure groups. On the contrary, selective activation of microglia was observed in the low blast group. In addition, increased astrocytes (GFAP), membrane signal transduction protein (Map2k1) and calcium regulator mechanosensitive protein (Piezo 2) were observed in the moderate blast group. Results from gene expression analysis suggested ongoing neuroprotection, as brain-derived neurotrophic factor (BDNF), glial-derived neurotrophic factor (GDNF) and Mn and CuZn superoxide dismutases (SOD) all increased in the low and moderate blast groups. Ongoing neuroprotection was further supported by increased SOD levels observed in the moderate group using immunohistochemistry. The gene expression level of glutamate aspartate transporter (GLAST) was upregulated in the low, but downregulated in the high blast group, while no changes were found in the moderate group. Overall, the data shown here provides evidence of a diverse neuroprotective and glial response to various levels of blast exposure. This mechanistic role of neuroprotection is vital in understanding ongoing cellular stress, both at the gene and protein levels, in order to develop interventional studies for the prognosis of injury.


The inhibitory effects of biomimetically designed peptides on α-synuclein aggregation.
Parkinson's disease is characterized by accumulation of inclusion bodies in dopaminergic neurons, where insoluble and fibrillar α-synuclein makes up the major component of these inclusion bodies. So far, several strategies have been applied in order to suppress α-synuclein aggregation and toxicity in Parkinson's disease. In the present study, a new database has been established by segmentation of all the proteins deposited in protein Data Bank. The database data base was searched for the sequences which adopt β structure and are identical or very similar to the regions of α-synuclein which are involved in aggregation. The adjacent β strands of the found sequences were chosen as the peptide inhibitors of α-synuclein aggregation. Two of the predicted peptides, namely KISVRV and GQTYVLPG, were experimentally proved to be efficient in suppressing aggregation of α-synuclein in vitro. Moreover, KISVRV exhibited the ability to disrupt oligomers of α-syn which are assumed to be the pathogenic species in Parkinson's disease.


Humoral and Cellular Immune Responses against SARS-CoV-2 after Third Dose BNT162b2 following Double-Dose Vaccination with BNT162b2 versus ChAdOx1 in Patients with Cancer.
Patients with cancer display reduced humoral responses after double-dose COVID-19 vaccination, whereas their cellular response is more comparable with that in healthy individuals. Recent studies demonstrated that a third vaccination dose boosts these immune responses, both in healthy people and patients with cancer. Because of the availability of many different COVID-19 vaccines, many people have been boosted with a different vaccine from the one used for double-dose vaccination. Data on such alternative vaccination schedules are scarce. This prospective study compares a third dose of BNT162b2 after double-dose BNT162b2 (homologous) versus ChAdOx1 (heterologous) vaccination in patients with cancer. A total of 442 subjects (315 patients and 127 healthy) received a third dose of BNT162b2 (230 homologous vs. 212 heterologous). Vaccine-induced adverse events (AE) were captured up to 7 days after vaccination. Humoral immunity was assessed by SARS-CoV-2 anti-S1 IgG antibody levels and SARS-CoV-2 50% neutralization titers (NT50) against Wuhan and BA.1 Omicron strains. Cellular immunity was examined by analyzing CD4+ and CD8+ T-cell responses against SARS-CoV-2-specific S1 and S2 peptides. Local AEs were more common after heterologous boosting. SARS-CoV-2 anti-S1 IgG antibody levels did not differ significantly between homologous and heterologous boosted subjects [GMT 1,755.90 BAU/mL (95% CI, 1,276.95-2,414.48) vs. 1,495.82 BAU/mL (95% CI, 1,131.48-1,977.46)]. However, homologous-boosted subjects show significantly higher NT50 values against BA.1 Omicron. Subjects receiving heterologous boosting demonstrated increased spike-specific CD8+ T cells, including higher IFNγ and TNFα levels. In patients with cancer who received double-dose ChAdOx1, a third heterologous dose of BNT162b2 was able to close the gap in antibody response.


Exploring satisfaction among women having a First Nations baby at one of three maternity hospitals offering culturally specific continuity of midwife care in Victoria, Australia: A cross-sectional survey.
Continuity of midwife care is recommended to redress the inequitable perinatal outcomes experienced by Aboriginal and Torres Strait Islander (First Nations) mothers and babies, however more evidence is needed about First Nations women's views and experiences of their care. This study aimed to explore levels of satisfaction among women having a First Nations baby, who received maternity care at one of three maternity services, where new culturally specific midwife continuity models had been recently implemented. Women having a First Nations baby who were booked for care at one of three study sites in Naarm (Melbourne), Victoria, were invited to complete one questionnaire during pregnancy and then a follow up questionnaire, 3 months after the birth. Follow up questionnaires were completed by 213 women, of whom 186 had received continuity of midwife care. Most women rated their pregnancy (80 %) and labour and birth care (81 %) highly ('6 or '7' on a scale of 1-7). Women felt informed, that they had an active say in decisions, that their concerns were taken seriously, and that the midwives were kind, understanding and there when needed. Ratings of inpatient postnatal care were lower (62 %), than care at home (87 %). Women having a First Nations baby at one of three maternity services, where culturally specific, continuity of midwife care models were implemented reported high levels of satisfaction with care. It is recommended that these programs are upscaled, implemented and sustained.


Testosterone replacement therapy for male hypogonadism: part III. Pharmacologic and clinical profiles, monitoring, safety issues, and potential future agents.
Male hypogonadism is associated with potentially distressing adverse effects on diverse organs and tissues. These include sexual dysfunction, particularly diminished libido, as well as mood disturbances, reduced lean body mass, and increased adipose-tissue mass. A wide range of effective and well-tolerated options exists. These include relatively noninvasive therapies, such as testosterone (T) gels and T patches; slightly more invasive treatments, such as the T buccal system; and invasive therapies, such as intramuscular T injections and subcutaneous depot implants (T pellets). Testosterone replacement therapy (TRT) can be individualized to enhance patient health and well-being. Screening and ongoing monitoring are necessary to ensure both the efficacy and safety of TRT, particularly prostate safety. Investigational agents, including selective androgen receptor modulators, may offer new pharmacodynamic and/or pharmacokinetic properties that enhance outcomes of TRT.


Sex hormones and female homosexuality: a critical examination.
To ascertain the validity of hormonal theories of human homosexuality, which are based on animal research, this article reviews psychoendocrine data on lesbian and transsexual women. Sex hormone levels were found to be normal in the majority of homosexual women, but about a third of the subjects studied had elevated androgen levels. In women with prenatal androgen excess, heterosexuality appears to be more frequent than bisexuality, and exclusive homosexuality is rare. Two recent reports suggest abnormalities of the neuroendocrine regulation of LH secretion in female transsexuals. Clearly, prenatal or postpubertal hormone levels do not determine the development of sexual orientation, but a facilitating neuroendocrine predisposition cannot be ruled out at present.


Outbreak of ertapenem-resistant Enterobacter cloacae urinary tract infections due to a contaminated ureteroscope.
Outbreaks of urinary tract infections (UTIs) due to contaminated ureteroscopes have been rarely reported. To report such an outbreak at a regional teaching hospital in southern Taiwan. From October to December 2010, ertapenem-resistant Enterobacter cloacae were identified from urine cultures of 15 patients who had undergone ureteroscopy prior to the infection. Three batches of surveillance cultures were obtained from the environmental objects and healthcare workers related to the procedures. Pulsed-field gel electrophoresis (PFGE) was used for bacterial typing. Antimicrobial susceptibility was assessed by disc diffusion and E-test methods. Polymerase chain reaction and sequencing were used to analyse β-lactamase genes. A total of 70 specimens were obtained during the first surveillance operation. One ertapenem-resistant E. cloacae was isolated from a ureteroscope. Although the disinfection protocols for ureteroscopes were revised and implemented, seven additional UTI cases were identified thereafter. The pathogen was identified from two subsequent surveillance cultures and was not eliminated until ethylene oxide sterilization was added to the disinfection protocol. PFGE revealed that all 15 isolates from the patients and the three isolates from the ureteroscope shared a common pattern with minor variance. Most isolates were resistant to gentamicin, levofloxacin, ceftriaxone, ceftazidime, and ertapenem. All isolates were susceptible to amikacin, imipenem, and meropenem. SHV-12 and IMP-8 genes were simultaneously identified in 16 of the 18 isolates. The outbreak of ertapenem-resistant E. cloacae was caused by a contaminated ureteroscope and was terminated by the implementation of a revised disinfection protocol for ureteroscopes.


Executive functions as mediators between socioeconomic status and academic performance in Chinese school-aged children.
It is well-documented that socioeconomic status (SES) and academic performance in school-aged children are closely related. However, little is known about how the three core executive functions (EFs), inhibition, working memory, and cognitive flexibility, mediate the association between the two. Moreover, most previous studies examined SES disparities in Western countries, how such disparities in EF and academic performance manifest in the Chinese context, where a distinctive EF profile and learning experience are observed, remains uncertain. The current study explored: (1) the mediating effects of the three core EFs in the association between SES and academic performance; and (2) the differences in EF and academic performance in three core subjects between Chinese children who are below and above the poverty line. Of the 385 students sampled, 205 are in the low-SES group and 180 are in the middle-high SES group. A structural equation model showed that the SES-academic performance relationship was fully mediated by cognitive flexibility and working memory but not inhibition. Working memory was a much stronger mediator than cognitive flexibility, suggesting that working memory may correlate with childhood SES and academic performance in Chinese children. An analysis of covariance suggested that compared to the middle-high SES group, the low-SES group demonstrated poorer working memory and academic performance in all three subjects after controlling for age and IQ. Interestingly, children with low-SES were found to have better cognitive flexibility than children with middle-high SES. These findings suggest that interventions targeting working memory may be an important area to improve children's academic performance.


Single MoO3 nanoribbon waveguides: good building blocks as elements and interconnects for nanophotonic applications.
Exploring new nanowaveguide materials and structures is of great scientific interest and technological significance for optical and photonic applications. In this work, high-quality single-crystal MoO3 nanoribbons (NRs) are synthesized and used for optical guiding. External light sources are efficiently launched into the single MoO3 NRs using silica fiber tapers. It is found that single MoO3 NRs are as good nanowaveguides with loss optical losses (typically less than 0.1 dB/μm) and broadband optical guiding in the visible/near-infrared region. Single MoO3 NRs have good Raman gains that are comparable to those of semiconductor nanowaveguides, but the second harmonic generation efficiencies are about 4 orders less than those of semiconductor nanowaveguides. And also no any third-order nonlinear optical effects are observed at high pump power. A hybrid Fabry-Pérot cavity containing an active CdSe nanowire and a passive MoO3 NR is also demonstrated, and the ability of coupling light from other active nanostructures and fluorescent liquid solutions has been further demonstrated. These optical properties make single MoO3 NRs attractive building blocks as elements and interconnects in miniaturized photonic circuitries and devices.


Metazoan evolution: some animals are more equal than others.
Comparison of newly available sequence data facilitates reconstruction of the gene inventory of the Urbilateria, the last common ancestor of flies, nematodes and humans. The most surprising outcome is that human genes seem to be closer to the bilaterian roots than previously assumed.


Monitoring carcinoembryonic antigen in colorectal cancer: is it still useful?
The results of a study conducted to determine the usefulness of carcinoembryonic antigen (CEA) monitoring in the follow-up of patients with resected colorectal cancer are reported herein. The subjects of this study were 125 patients in whom CEA had been determined preoperatively and 239 patients in whom CEA had been monitored postoperatively. The results revealed increased preoperative CEA in only 24% of the subjects, and that this increment was correlated with subsequent more advanced tumor stage and a higher recurrence rate (P < 0.01). The postoperative CEA level exceeded the threshold in 71% of the patients affected by recurrence, 94.4% of whom developed liver metastases and 50%, nonhepatic recurrence. This marker showed elevated sensitivity for liver metastases (99%), whereas the sensitivity was lower for nonhepatic recurrence of the disease (94%). Thus, we concluded that CEA monitoring can be useful for preoperative colorectal tumor grading, even if its validity in the early diagnosis of recurrence is problematic, especially in terms of radical repeated surgery and survival.


What constitutes a neurosurgical emergency?
Because of the large numbers of head injuries, physicians are frequently called upon to evaluate and initiate treatment in these acute emergencies. Deterioration of such patients results from direct injury to the brain tissue and a subsequent increase in the intracranial pressure. The pathophysiology of brain injury is manifested in a cycle of detrimental events with increased tissue damage and progressive neurological deficit. Current concepts in management of head injuries aimed at interrupting this cycle of events are discussed in detail.


"I think to myself 'why now?'" - a qualitative study about endometriosis and pain in Austria.
Endometriosis is a chronic, benign, and oestrogen-dependent condition and about 10-15% of all women of reproductive age are affected by endometriosis worldwide. It is not curable and pain is one of the most common symptoms of endometriosis and leads to low quality of life in affected women. To our knowledge, in German-speaking countries, no studies with qualitative methods approaches are available concerning women who suffer from pain caused by endometriosis and possible associated coping strategies. Our study aims to familiarise ourselves with the individual pain experience of selected women who suffer from endometriosis in Austria and their coping strategies. A qualitative study design was based on problem-centred interviews for data collection and qualitative content analysis for data analysis. The research participants were women aged between 18 and 55 diagnosed with endometriosis and living in Austria. The interview period was from 27 February to 26 March 2019 and interviews lasted between 50 and 75 min. Eight categories were formulated, of which category 3 (thoughts and feelings regarding endometriosis and pain - 'why?'), category 5 (effects and changes caused by endometriosis and pain - 'quality of life'), category 7 (taboos - 'don`t talk about it'), and category 8 (talking about it - 'contact with others in the same position') were relevant for this article. The remaining four categories [1-4] have already been published elsewhere. Our data show that the social environment plays a fundamental role in coping strategies concerning pain caused by endometriosis. Women in our study reported that exchange with peers offers support. This opens a door for information events, patient organizations like support groups, and the inclusion of these in the supporting system. Involving occupational medicine and workplace health promotion departments in companies should be further goals to support affected women.


Application of alpha7 nicotinic acetylcholine receptor agonists in inflammatory diseases: an overview.
Inflammatory disorders are characterized by the influx of immune cells into the vascular wall of veins and/or arteries in response to stimuli such as oxidized-LDL and various pathogens. These factors stimulate the local production of pro-inflammatory cytokines by macrophages and other cells that promote various inflammatory diseases such as atherosclerosis, Crohn's, Alzheimer's and diabetes. Numerous cytokines play a significant role in this process, though tumor necrosis factor (TNF) and various interleukins are thought to be among the most important regulators. These proinflammatory cytokines promote the above-described diseases by inducing endothelial cell dysfunction. In this brief commentary we will discuss some of the latest advances and discoveries in the treatment of these inflammatory diseases, making use of alpha7 nicotinic acetylcholine receptor (alpha7 nAChR) agonists.


Behavior of N-methylated allopurinols and related 4-thioxopyrazolo [3,4-d]pyrimidines towards bovine milk xanthine oxidase.
1. All available N-mono- and N,N'-dimethylallopurinols and the corresponding 4-thioxo derivatives have been tested as substrates or inhibitors of bovine milk xanthine oxidase (xanthine: oxygen oxidoreductase, EC 188.8.131.52). 2. None of the compounds tested revealed any inhibitory activity towards the enzyme. 3. All compounds were resistant to enzymic oxidation, with the exception of 7-methylallopurinol and its 4-thioxo analog. Both these compounds were attacked at position 6. 7-Methylallopurinol was oxidised nearly ten times faster than the isomeric 3-methylhypoxanthine. 4. These observations can be explained by assuming that for attack at C-6, the enzyme must bind both to N-1 and N-2 in the pyrazole ring and causes tautomerisation, which places a double bond at position 5,6 in the pyrimidine ring. This activation process resembles the activation of hypoxanthine.


Thresholds for perceiving metallic taste at high magnetic field.
To perform an initial characterization of the metallic taste effect observed by some workers when moving around an MRI scanner. A total of 21 subjects performed controlled movements in the stray field of a 7-T scanner. Rates of change of magnetic flux were recorded during the study using a custom-built three-axis coil unit connected to a data logger. Relatively normal movements could generate switched fields of 2 T/second. Of the 21 subjects, 12 detected a metallic taste, but the threshold at which it was perceived varied greatly between subjects, with the minimum dB/dt value at which such a taste was detected being 1.3 T/second. The threshold also depended on the direction of movement. This study indicates that 50% of subjects will perceive a metallic taste for head shaking with a period of 1.5 seconds (magnetic field in an anterior/posterior direction) causing a dB/dt of 2.3 +/- 0.3 T/second. The presence of dental fillings is not a requirement for the sensation of metallic taste.


Effects of continuous positive airway pressure on pulmonary function and exercise tolerance in patients with congestive heart failure.
Continuous positive airway pressure (CPAP) has been used to improve cardiopulmonary function and reduce pulmonary edema symptoms in patients with congestive heart failure (CHF). The objective of this study was to evaluate the efficacy of CPAP therapy on pulmonary function and exercise tolerance in patients with CHF. Prospective blind randomized clinical study. Twenty-four patients with class II or III CHF and dilated cardiomyopathy were randomly assigned to 30 min of CPAP therapy and respiratory exercises (CPAP group) or respiratory exercise only (control group) once a day for 14 days. Evaluation of pulmonary function was performed measuring FEV1 and FVC. Exercise tolerance was assessed measuring the distance walked during the 6-min walking test (6MWT). These parameters were measured before treatment and 4 days, 9 days, and 14 days later. CPAP therapy caused a progressive increase (p < 0.05) in both FVC (maximum of 16% after 9 days) and FEV1 (maximum of 14% after 14 days) compared to basal values, without significant changes in the control group. The 6MWT showed a progressive improvement in the distance walked in the CPAP group, reaching approximately 28% above the basal values in the CPAP group and without significant changes in the control group. These data show that the use of CPAP therapy for 2 weeks on a daily basis is able to enhance pulmonary function and consequently improve the tolerance to physical activities in patients with CHF. The clinical implication of this finding is that CPAP therapy could potentially be used as an adjunct to the treatment of CHF patients.


The nitric oxide donor sodium nitroprusside protects against hepatic microcirculatory dysfunction in early endotoxaemia.
Endotoxin rapidly inhibits the activity of the constitutive endothelial nitric oxide synthase (ecNOS); this precedes the production of NO from inducible NOS (iNOS). This leaves a period in early endotoxaemia with a supposed scarcity of NO. The present study was conducted to examine the effects of external supplementation of NO on liver microcirculation and function. 13 male Sprague Dawley rats. The rats underwent laparotomy, and the left liver lobe was exteriorised. All animals were given a bolus dose of endotoxin (LPS) 5 mg/kg intraportally. One group (n = 6) had a continuous infusion of sodium nitroprusside (SNP) 1.4 microg/kg per min started concurrently, the other group (n = 7) was treated with normal saline. The study was terminated after 3 h LPS. Intravital microscopy was performed at baseline, at 2 h and 3 h LPS. Hepatic function was assessed by arterial ketone body ratio, acid base values, and bile flow. At baseline 1% of the sinusoids were without perfusion. After 2 h LPS this figure had risen to 9.8+/-1.5% in the SNP group versus 16.9+/-1.4% in the controls (p < 0.05 vs controls). The corresponding values after 3 h LPS were 13.5+/-1.5 versus 19.3+/-1.5% (p < 0.05 vs controls). The leukocyte count in sinusoids and venules had a similar development. Functional parameters were all slightly better preserved in the SNP group, but with no individual significance versus controls. Infusion of the NO donor SNP in early endotoxaemia attenuates the detrimental effects of LPS on liver microcirculation, most probably by alleviating a relative deficit of NO at the microcirculatory level.


Mediating and moderating effects of stress perception and situation type on coping responses in women with disordered eating.
Effects of stress perception and situation type on coping behavior were assessed in undergraduate females. Women with high (n=28) and low (n=28) Eating Attitudes Test (EAT) scores listened to audiotaped scenes representing two different situation types and two levels of stress. As predicted, the High EAT group rated scenes as more stressful and endorsed more of the maladaptive emotion-oriented coping, compared to the low EAT condition. Importantly, analyses revealed that stress perception mediated the relationship between group membership and emotion-oriented coping. Scene type appeared to moderate the effect of group on emotion-oriented coping. However, stress perception was shown to mediate, or account for, the moderating effect of situation type; a relationship termed mediated moderation. So although the two groups appeared to differ in the effect of situation type on coping, this apparent interaction effect can be attributed to differences in their perception of stress. Finally, women with High EAT scores did not exhibit a deficit in the more productive task-oriented coping behaviors, nor did they exhibit an avoidant coping style. In sum, the maladaptive coping used by women with disordered eating may represent characteristic emotional responses to exaggerated perceptions of stress and a preoccupation with issues specific to body image and food, rather than deficits in productive coping skills or an avoidant coping style.


Impact of letrozole supplementation during ovarian stimulation for fertility preservation in breast cancer patients.
Oocyte and/or embryo vitrification after controlled ovarian stimulation (COS) represents the most established method of fertility preservation (FP) before cancer treatment. However, traditional COS regimens are associated with supraphysiologic serum estradiol and are therefore not recommended in estrogen-sensitive diseases such as breast cancer (BC). To protect the patients from the potential deleterious effects of elevated estrogen levels during COS for FP, protocols using aromatase inhibitors (letrozole) were developed. The present study aims at investigating whether COS with letrozole supplementation (COSTLES) modifies ovarian response in BC patients. One hundred and seventy-seven BC patients candidates for FP using oocyte and/or embryo vitrification following COS referred to our center between July 2013 and December 2016 were included in this retrospective case-control study. 94 patients underwent COSTLES while 83 had standard GnRH antagonist protocol. The number of oocytes retrieved, oocyte maturation rates, number of oocytes vitrified and follicle responsiveness to FSH assessed by the Follicular Output Rate (FORT) were assessed. Women in both groups were comparable in terms of age and ovarian reserve tests leading to a similar number of oocyte recovered (13.1 ± 10.0 vs. 12.2 ± 8.0 oocytes, respectively, NS). However, oocyte maturation rates were significantly lower in COSTLES compared to standard protocol (64.9 ± 22.8 vs. 77.4 ± 19.3%, p < 0.001). As a result, the number of mature oocyte vitrified was lower in COSTLES group (7.8 ± 5.3 vs. 10.3 ± 8.5 oocytes, p < 0.001 respectively). Despite similar response to exogenous FSH, BC patients having undergone COSTLES show reduced oocyte maturation rates in comparison with those having received standard stimulation regimen.


[Randomized experimental study to evaluate the effectiveness of the Flebitis Zero project in Navarre].
Phlebitis is one of the most frequent complications in peripheral venous catheters. The aim was to compare the frequency of phlebitis before and after the implementation of the Flebitis Zero project, in order to ascertain its effectiveness and what factors were associated with this complication. Randomized pre-post intervention (Flebitis Zero project) study performed in the Hospital Universitario de Navarra (Pamplona, Spain) between 2015 and 2019. The intervention consisted of training of professionals along with the implementation of measures in order to lower the incidence of complications associated with the peripheral venous catheters. The control group (CG) was selected prior the intervention and the Intervention Group (IG) in the subsequent five years. The study included 656 patients, 96 (14.6%) in the CG and 569 in the IG. Lower frequency of complications (49.0 vs 37.1%; p<0.001), especially accidental PVC exit (18.8 vs 8.1%), and of phlebitis (13.5 vs 11.8%). The intervention and the use of fully reinforced dressings were associated to happening of phlebitis. The implantation of the Flebitis Zero project prevented the occurrence of phlebitis (OR = 0.78; IC95%: 0.42-0.86; p = 0.042). The implementation of the measures used in the Phlebitis Zero project was associated with a significant reduction of the complications linked to the use of the peripheral venous catheters; the frequency of phlebitis dropped, although nowhere near the recommended level.


Relationship between clinical data and gene expression in the HER2/ErbB2-dependent signaling pathway in patients with acute heart failure.
Anticancer treatment with the human epidermal growth factor receptor (HER) 2 inhibitors can lead to significant myocardial dysfunction. The primary aim of the study was to estimate the possible association between gene expression in the ErbB signaling pathway and selected clinical event data in patients with acute heart failure. Twenty-four patients (19 males), aged 68.6 ± 12.3 years, were diagnosed and treated due to acute heart failure. The globaltest method was used for the correlation between blood nuclear cells' gene expression in the ErbB pathway (KEGG pathway id 04012) and important clinical data. Decreased expression of ErbB2/HER2 was found to be associated with the release of troponin and the need for inotropic support, whereas decreased neuregulin 1 (NRG1) expression was found to be associated with a decrease of ejection fraction below 40 % (globaltest p-value < 0.05). In summary, the ErbB signaling pathway and, especially, HER2/ErbB2 receptor expression are significantly associated with some of the recognized, clinically significant parameters of patients with acute heart failure. Evaluation of the molecular function of the HER2 receptor may be essential for the prognosis and targeted therapy of heart diseases.


How to avoid esophageal perforation while performing laparoscopic dissection of the hiatus.
An increasing number of surgeons attempt advanced laparoscopic procedures, involving the distal esophagus such as Nissen fundoplication, truncal vagotomy, and Heller's myotomy. At this time, there are probably as many techniques as there are surgeons. The authors have tried to provide a "ready to use" universal strategy that details how to approach the distal esophagus while avoiding the dangerous pitfalls of surgery in that area.


Genotypic evolution and antigenicity of H9N2 influenza viruses in Shanghai, China.
H9N2 influenza viruses have been circulating in China since 1994, but a systematic investigation of H9N2 in Shanghai has not previously been undertaken. Here, using 14 viruses we isolated from poultry and pigs in Shanghai during 2002 and 2006-2014, together with the commercial vaccine A/chicken/Shanghai/F/1998 (Ck/SH/F/98), we analyzed the evolution of H9N2 influenza viruses in Shanghai and showed that all 14 isolates originated from Ck/SH/F/98 antigenically. We evaluated the immune protection efficiency of the vaccine. Our findings demonstrate that H9N2 viruses in Shanghai have undergone extensive reassortment. Various genotypes emerged in 2002, 2006 and 2007, while during 2009-2014 only one genotype was found. Four antigenic groups, A-D, could be identified among the 14 isolates and a variety of antigenically distinct H9N2-virus-derived avian influenza viruses (AIVs) circulated simultaneously in Shanghai during this period. Challenge experiments using vaccinated chickens indicated that the vaccine prevented shedding of antigenic group A and B viruses, but not those of the more recent groups C and D. Genetic analysis showed that compared to the vaccine strain, representative viruses of antigenic groups C and D possess greater numbers of amino acid substitutions in the hemagglutinin (HA) protein than viruses in antigenic groups A and B. Many of these substitutions are located in antigenic sites. Our results indicate that the persistence of H9N2 AIV in China might be due to incomplete vaccine protection and that the avian influenza vaccine should be regularly evaluated and updated to maintain optimal protection.


Building leadership capacity through peer career coaching: a case study.
Today's demanding healthcare environment requires resiliency, creativity and innovation in delivery of patient care and service. Hospitals must create a workplace where staff are supported to develop professionally as knowledge workers. In 2003, University Health Network (UHN) partnered with donnerwheeler, career planning and development consultants, to provide a program for its 2,700 registered nurses. One component of this project, a peer coaching program called Coach Mastery, is profiled in this case study, which describes how it was implemented and the successes, challenges and outcomes in building internal leadership capacity and supporting staff development through career planning and development.


Long-term results of corneal transplantations in keratoconus patients.
In this retrospective study all 86 corneal transplantations performed for keratoconus in the Rotterdam Eye Hospital between 1969 and 1978 were included. Sixty seven perforating and 19 lamellar corneal grafts had been performed. The average follow-up was 59 months. Immune reactions occurred 12 times in 11 eyes with a perforating corneal graft (16%); in no case did this lead to permanent opacities in the graft. The visual result of the perforating graft is in general better than that of the lamellar graft. A lamellar graft is indicated, in our opinion, when the risk of trauma is high and the opportunities for follow-up insufficient, and when the peripheral cornea is markedly thin. In most cases we prefer a perforating graft because the results are better.


Search for circulating immune complexes and activation of the complement system in relation to estrogen treatment.
By employing an ammonium sulphate precipitation technique, previous studies have shown circulating immune complexes (cIC) in an increased proportion of women using oral contraceptives, as compared with non-users. The pathogenicity of cIC is well established, as deposition in organs may lead to activation of the complement system and thereby inflammation and tissue destruction. The purpose of the present study was to determine whether estrogen treatment could induce the formation of cIC, as measured by more specific immune complex assays. To test for complement activation, plasma samples were analysed for the complement split product C3d. Thirty-one women treated with estrogen for menopausal hormone deficiency symptoms and 38 untreated controls were analysed for cIC. Using a solid-phase C1q-anti-IgG binding assay, the mean level of cIC in the estrogen-treated group did not differ significantly from that of the untreated control group (p greater than 0.05). In a solid-phase C1q-protein A binding assay a very small average increase in the cIC values for the estrogen-treated group in comparison with the untreated group was found, expressing a significant difference (p = 0.04). Moreover, the C3d levels for the estrogen-treated group did not differ from the untreated control group. Finally, no differences could be demonstrated in cIC and C3d levels between patients treated with synthetic estrogens and patients treated with natural estrogens.


Clinical role of granulocyte-macrophage colony-stimulating factor.
The activity of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) for increasing leukocyte production and enhancing mature neutrophil function has been clearly demonstrated in phase I clinical trials in patients with a variety of hematologic and malignant diseases. The focus of current studies includes determination of optimal administration schedules and its use in combination with myelosuppressive chemotherapy, radiation therapy or antimicrobial agents. The utility of GM-CSF as a stimulant of host defense against infections and tumors is also under study.


Single-Cell Chromatin and Gene-Regulatory Dynamics of Mouse Nephron Progenitors.
We reasoned that unraveling the dynamic changes in accessibility of genomic regulatory elements and gene expression at single-cell resolution will inform the basic mechanisms of nephrogenesis. We performed single-cell ATAC-seq and RNA-seq both individually (singleomes; Six2GFP cells) and jointly in the same cells (multiomes; kidneys) to generate integrated chromatin and transcriptional maps in mouse embryonic and neonatal nephron progenitor cells. We demonstrate that singleomes and multiomes are comparable in assigning most cell states, identification of new cell type markers, and defining the transcription factors driving cell identity. However, multiomes are more precise in defining the progenitor population. Multiomes identified a "pioneer" bHLH/Fox motif signature in nephron progenitor cells. Moreover, we identified a subset of Fox factors exhibiting high chromatin activity in podocytes. One of these Fox factors, Foxp1, is important for nephrogenesis. Key nephrogenic factors are distinguished by strong correlation between linked gene regulatory elements and gene expression. Mapping the regulatory landscape at single-cell resolution informs the regulatory hierarchy of nephrogenesis. Paired single-cell epigenomes and transcriptomes of nephron progenitors should provide a foundation to understand prenatal programming, regeneration after injury, and ex vivo nephrogenesis.


The significance of insecticide-resistant strains; with special reference to pests of medical importance.
The author discusses the meaning of insecticide resistance and the manner in which it should be detected and measured. From some recent data he gives a number of examples of measurements of resistance in mosquitos of different species in various areas. He then proceeds to a speculative discussion on the way in which insecticide resistance arises, whether it can be prevented and overcome, its importance in the past and at present, and finally its future prospects.


Evaluation of DNA damage and physiological responses in Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) exposed to sub-lethal diclofenac (DCF).
The frequent bioaccumulation of pharmaceuticals in the aquatic ecosystem has raised a concern about their possible ecotoxicological consequences. DNA damage, haematological changes and activities of oxidative stress enzymes in Nile tilapia, Oreochromis niloticus in response to diclofenac (DCF) exposure were investigated for up to 60 days at the concentrations of 0.17, 0.34 and 0.68mgL-1 in the fish liver. Evaluation of genotoxic effects of the drug in the liver, using single-cell gel electrophoresis, showed DNA damage on exposure at the concentrations of 0.34 and 0.68mgL-1 after day 30. Compared with the control, there was a reduction in haemoglobin and red blood cell counts with a significant increase (p<0.05) in white blood cell counts, mean corpuscular volume and mean corpuscular haemoglobin level after day 30 at 0.34 and 0.68mgL-1. The levels of pack cell volume, red cell distribution width and mean corpuscular haemoglobin concentration were not significant (p>0.05) between the exposed group and the control. The indices of hepatic oxidative stress biomarkers, including lipid peroxidation and carbonyl protein, showed elevated level, depicting a positive correlation with both time and concentration. More so, activity of catalase was inhibited while reduced glutathione level decreased in the liver tissue. There was increase in the activities of superoxide dismutase, glutathione peroxidase and glutathione-S-transferase after 30 days at 0.34mgL-1. Further, activity of Na+-K+-ATPase in the tissue was significantly inhibited (p<0.05) at the end of 60 days. Prolonged exposure to diclofenac at sub-lethal concentration can cause both DNA and oxidative damages in O. niloticus, suggesting the use of oxidative stress biomarkers as early warning signals in environmental monitoring of residual pharmaceutical and assessment.


Purification of bovine liver S-adenosylhomocysteine hydrolase by affinity chromatography on blue dextran-agarose.
S-Adenosylhomocysteine (AdoHcy) hydrolase (adenosylhomocysteinase, EC 126.96.36.199) was purified from bovine liver by conventional protein purification procedures (differential centrifugation, ammonium sulfate fractionation and DEAE-cellulose chromatography) followed by affinity chromatography on blue dextran coupled to agarose. The enzyme was eluted from the blue dextran-agarose column with adenosine and the adenosine was removed by chromatography on Sephadex G-75. The affinity chromatography step resulted in a substantial increase in total AdoHcy hydrolase activity (about 600%) suggesting either removal of some inhibitory substance or a change in the structure of the protein producing a more catalytically efficient enzyme. The isolation procedure afforded over 3400-fold purification of the enzyme, which was shown to be homogeneous by polyacrylamide gel electrophoresis. Using high pressure liquid chromatography, the nucleotide content of the freshly purified enzyme was determined to be 2 mol of nicotinamide adenine nucleotide per mol of enzyme tetramer. The ratio of the reduced to the oxidized form of the nucleotide was correlated to the activity of the enzyme preparation.


Fullerene (C60) immunoconjugates: interaction of water-soluble C60 derivatives with the murine anti-gp240 melanoma antibody.
The first fullerene (C60) immunoconjugates have been prepared and characterized as an initial step toward the development of fullerene immunotherapy (FIT).


Assessment of right ventricular function postretrograde cardioplegia by transesophageal echocardiography.
The impact of continuous retrograde cardioplegia (RCP) on right ventricular (RV) function was evaluated prospectively with intraoperative transesophageal echocardiography (TEE) in 36 patients (23 males, 13 females) with a mean age of 60.4 years (ages 24-82). Operative procedures included 12 aortic valve replacements, 16 mitral valve repair/replacements, both with or without an associated cardiac operation, and 8 Ross procedures. Mean cardiopulmonary bypass (CPB) time was 123.3 minutes (66-280 minutes) with an average cross-clamp time of 88.9 minutes (43-199 minutes). The amount of cold blood RCP ranged from 3160-18,500 mL (mean = 7382.5), and the average pulmonary artery pressure was 35/18 mmHg. The coronary sinus was distally snared in 11 patients. TEE documented biventricular global dysfunction in two patients and post-CPB with preservation of the left ventricular function in all other patients. Localized akinesis of the RV apex and outflow tract were noted in three patients, and isolated worsening tricuspid insufficiency of moderate to severe intensity in six patients. Two of the six patients with worsening tricuspid insufficiency belonged to the snared coronary sinus group (11 patients). All of the documented RV dysfunctions were new and showed no correlation with the perfusion data. (1) RV apex, RV outflow tract and tricuspid valve were particularly subject to important dysfunction post-RCP; (2) RCP did not protect RV adequately in 11 patients (31%); and (3) TEE is a convenient intraoperative technique in evaluating RV dysfunction.


Preferences for descriptors of hypnosis: a brief communication.
Alternative descriptors of the capacity to experience hypnosis, intended to describe the same phenomenon, appear in the current literature. Published members of the Society for Clinical and Experimental Hypnosis (SCEH) were surveyed to determine their preferences. The descriptors were empirically derived from recent International Journal of Clinical and Experimental Hypnosis articles and input from the executive committee of SCEH. Participants also indicated their primary theoretical conceptualization of hypnosis. Hypnotizability was chosen nearly 4 times more frequently than the next most favored choice (susceptibility) as a descriptor of hypnotic talent. Hypnosis as an "identifiable state" was chosen more than 4 times more frequently than the socio-cognitive version. This latter finding suggests that the notion of the continued debatability of hypnosis as primarily a state is now shared by only a few.


The first crystal structure of a family 31 carbohydrate-binding module with affinity to beta-1,3-xylan.
Here, we present the crystal structure of the family 31 carbohydrate-binding module (CBM) of beta-1,3-xylanase from Alcaligenes sp. strain XY-234 (AlcCBM31) determined at a resolution of 1.25A. The AlcCBM31 shows affinity with only beta-1,3-xylan. The AlcCBM31 molecule makes a beta-sandwich structure composed of eight beta-strands with a typical immunoglobulin fold and contains two intra-molecular disulfide bonds. The folding topology of AlcCBM31 differs from that of the large majority of other CBMs, in which eight beta-strands comprise a beta-sandwich structure with a typical jelly-roll fold. AlcCBM31 shows structural similarity with CBM structures of family 34 and family 9, which also adopt structures based on immunoglobulin folds.


Cubic sub-20 nm NaLuF(4)-based upconversion nanophosphors for high-contrast bioimaging in different animal species.
A new upconversion luminescence (UCL) nanophosphors based on host matrix of cubic NaLuF(4) with bright luminescence have been synthesized by a solvothermal method, facilitate the nanocrystals potential candidates for imaging in vivo, especially large-animals. The sub-20 nm NaLuF(4) co-doped Yb(3+) and Er(3+) (Tm(3+)) showed about 10-fold stronger UCL emission than that of corresponding hexagonal NaYF(4)-based nanocrystals with a 20 nm diameter. Near-infrared to near-infrared (NIR-to-NIR) UCL emission of PAA-coated NaLuF(4):20%Yb,1%Tm (PAA-Lu(Tm)) can penetrate >1.5 cm tissue of pork with high contrast. Based on super-strong UCL emission and deep penetration, PAA-Lu(Tm) as optical bioprobe has been demonstrated by in vivo UCL imaging of a normal black mouse, even rabbit with excellent signal-to-noise ratio. Furthermore, such cubic NaLuF(4)-based nanophosphor was applied in lymph node imaging of live Kunming mouse with rich white fur.


Feeding infants below 29 weeks' gestation with abnormal antenatal Doppler: analysis from a randomised trial.
To describe feeding and gastrointestinal outcomes in growth-restricted infants <29 weeks' gestation and to determine the rate of feed advancement which they tolerate. Analysis of prospectively collected data from a randomised feeding trial, the Abnormal Doppler Enteral Prescription Trial (ADEPT). 54 neonatal units in the UK and Ireland. 404 preterm, growth-restricted infants with abnormal antenatal Doppler studies from ADEPT. 83 infants <29 weeks and 312 infants ≥ 29 weeks' gestation were included in this analysis. In ADEPT, infants were randomised to start milk 'early' on day 2 after birth, or 'late' on day 6. Subsequent feed advancement followed a regimen, which should have achieved full feeds by day 16 in the early and day 20 in the late group. Full feeds were achieved later in infants <29 weeks; median age 28 days {IQR 22-40} compared with 19 days {IQR 17-23} in infants ≥ 29 weeks (HR 0.35, 95% CI 0.3 to 0.5). The incidence of necrotising enterocolitis was also higher in this group; 32/83 (39%) compared to 32/312 (10%) in those ≥ 29 weeks (RR 3.7, 95% CI 2.4 to 5.7). Infants <29 weeks tolerated very little milk for the first 10 days of life and reached full feeds 9 days later than predicted from the trial regimen. Growth-restricted infants born <29 weeks' gestation with abnormal antenatal Doppler failed to tolerate even the careful feeding regimen of ADEPT. A slower advancement of feeds may be required for these infants. ISRCTN87351483.


Assessment and Correlation of Variation in Lingual Cortical Plate Thickness with Different Angulations of Impacted Mandibular Third Molar Using Cone Beam Computed Tomography in North Indian Population.
The extraction of impacted mandibular third molar is associated with various types of intraoperative and postoperative complications, one of them is injury to lingual nerve. The present study aims to assess and correlate the variation in lingual cortical plate thickness with different angulations and determine the topographic relationship between the root apex of impacted mandibular third molar and lingual cortical plate using Cone Beam Computed Tomography (CBCT). This prospective cohort study enrolled 140 patients with impacted mandibular third molars who underwent preoperative CBCT imaging. The CBCT images were used to evaluate the outcome variables such as lingual cortical plate thickness and topographical relationship between root apex of impacted mandibular third molar and lingual cortical plate. The predictor variables included age, gender, inclination of third molar, mesiodistal angle, buccolingual angle and lingual plate morphology. The temporary lingual nerve paresthesia was reported by 1 patient (0.71%) out of 140 patients. The lingual bone in distoangular and vertical impacted third molar was found to be 1.20 times thicker than mesioangular and horizontal teeth. Lingual bone thinning at mid-root level was observed in maximum number of cases with mesioangular teeth (68.5%), whereas horizontal impacted teeth showed lingual bone thinning (90.9%) at root apex level. The mean buccolingual angle of impacted mandibular third molar was found to be significantly associated with lingual bone perforation (p value = 0.0258). The morphology of lingual plate was observed as undercut type (37.14%) followed by slanted (36.43%), parallel (19.29%) and round type (7.14%). Increase in buccolingual angle of impacted mandibular third molar decreases lingual bone thickness which is the natural barrier for protecting injury to lingual nerve. Also undercut and slanted lingual plate shapes were recognised as risk factors for contact/perforation between root apex and lingual plate. Therefore, proper screening and planning of high risk patients before third molar extraction is crucial.


Successful parallel development and integration of a plasmid-based biologic, container/closure system and electrokinetic delivery device.
We have developed three major technologies that allow plasmid-based products to be used for large-scale vaccination or therapeutic protein applications. Our team has integrated these components into one complete, cost-effective, easy-to-use system capable of rapid implementation under field conditions. The proprietary manufacturing process uses a lysis method and membrane-based chromatography to rapidly produce large-scale batches of plasmid. Plasmid doses are filled into the Becton-Dickinson Uniject container/closure system. The Uniject adapts to the electrode array of our constant current electrokinetic device, such that the plasmid is delivered in the area of tissue defined by the electrodes. Thus, plasmid uptake and expression levels are dramatically improved. This is the first completely integrated delivery system for plasmid-based products.


Self-Efficacy and Social Support are Associated with Disability for Ambulatory Prosthesis Users After Lower-Limb Amputation.
Interventions targeting psychosocial factors may improve rehabilitation outcomes for prosthesis users after lower-limb amputation (LLA), but there is a need to identify targeted factors for minimizing disability. To identify psychosocial factors related to disability for prosthesis users after LLA in middle age or later. Cross-sectional study. General community. Participants with LLA (N = 122) were included in this cross-sectional study if their most recent LLA was at least 1 year prior, they were ambulating independently with a prosthesis, and they were between 45 and 88 years old. Not applicable. Disability, the primary outcome, was measured using the World Health Organization Disability Assessment Schedule 2.0 (WHODAS). Candidate psychosocial variables included self-efficacy, social support, and motivation, measured using the Self-Efficacy of Managing Chronic Disease questionnaire (SEMCD), Multidimensional Scale of Perceived Social Support questionnaire (MSPSS), and modified contemplation ladder (mCL), respectively. The hypothesis was that greater self-efficacy, social support, and motivation would be associated with lower disability when controlling for covariates. The covariate model, including etiology, age, sex, U.S. military veteran status, LLA characteristics, time since LLA, medical complexity, and perceived functional capacity, explained 66.1% of disability variability (WHODAS 2.0). Backward elimination of candidate psychosocial variables stopped after removal of motivation (P = .10), with self-efficacy (P < .001) and social support (P = .002) variables remaining in the final model. The final model fit was statistically improved (P < .001) and explained an additional 6.1% of disability variability when compared to the covariate model. Greater self-efficacy and social support are related to lower disability after LLA. Findings suggest there may be a role for interventions targeting increased physical function, self-efficacy, and social support for ambulatory prosthesis users after LLA in middle age or later, especially when complicated by multiple chronic conditions.


A novel immobilization strategy using oligonucleotide as linker for small molecule microarrays construction.
A novel immobilization method based on oligonucleotide as linker has been developed for small molecule microarrays (SMMs) construction. The oligonucleotide tail was employed as a linker in solid-phase synthesis. Small molecules could be easily conjugated at the 5' end of the oligonucleotide, previously modified with a functional group. Being a reactive species, the oligonucleotide was activated by UV irradiation, for the attachment of the conjugate to the slide surface. The method was successfully applied to structurally distinct small molecules, including biotin, antibiotic and drug. This immobilization strategy showed high efficiency, 1.1fmol of small molecules in the spotting solution per spot gave a detectable signal (mean S/N=10.9). The results suggest that it is very promising for exploring interaction between small molecules and proteins, and high throughput detecting the chemical compounds.


Studies of DNA dumbbells. IV. Preparation and melting of a DNA dumbbell with the 16 base-pair sequence 5'G-T-A-T-C-C-C-T-C-T-G-G-A-T-A-C3' linked on the ends by dodecyl chains.
The preparation and melting of a 16 base-pair duplex DNA linked on both ends by C12H24 (dodecyl) chains is described. Absorbance vs temperature curves (optical melting curves) were measured for the dodecyl-linked molecule and the same duplex molecule linked on the ends instead by T4 loops. Optical melting curves of both molecules were measured in 25, 55, and 85 mM Na+ and revealed, regardless of [Na+], the duplex linked by dodecyl loops is more stable by at least 6 degrees C than the same duplex linked by T4 loops. Experimental curves in each salt environment were analyzed in terms of the two-state and multistate theoretical models. In the two-state, or van't Hoff analysis, the melting transition is assumed to occur in an all-or-none manner. Thus, the only possible states accessible to the molecule throughout the melting transition are the completely intact duplex and the completely melted duplex or minicircle. In the multistate analysis no assumptions regarding the melting transition are required and the statistical occurrence of every possible partially melted state of the duplex is explicitly considered. Results of the analysis revealed the melting transitions of both the dodecyl-linked molecule and the dumbbell with T4 end loops are essentially two state in 25 and 55 mM Na+. In contrast, significant deviations from two-state behavior were observed in 85 mM Na+. From our previously published melting data of DNA dumbbells with Tn end loops where n = 2, 3, 4, 6, 8, 10, 14 [T. M. Paner, M. Amaratunga, and A. S. Benight, (1992) Biopolymers, Vol. 32, pp. 881-892] and the dumbbell with T4 end loops of this study, a plot of d(Tm)/d ln [Na+] was constructed. Extrapolation of this data to n = 1 intersects with the value of d (Tm)/d ln [Na+] obtained for the alkyl-linked dumbbell, suggesting the salt-dependent stability of the alkyl-linked molecule behaves as though the duplex of this molecule were linked by end loops comprised of a single T residue.


A major locus on chromosome 3p22 conferring predisposition to human herpesvirus 8 infection.
Infection with human herpesvirus 8 (HHV-8), the etiological agent of Kaposi's sarcoma, has been shown to display strong familial aggregation, in countries in which HHV-8 infection is endemic. We investigated 40 large families (608 subjects aged one to 88 years) living in an isolated area of Cameroon in which HHV-8 is highly endemic. We performed a two-step genetic analysis for HHV-8 infection status (HHV-8+/HHV-8- determined by immunofluorescence) consisting of an initial segregation analysis followed by a model-based genome-wide linkage analysis. Overall HHV-8 seroprevalence was 60%, increasing with age. Segregation analysis provided strong evidence for a recessive major gene conferring predisposition to HHV-8 infection. This gene is predicted to have a major effect during childhood, with almost all homozygous predisposed subjects (∼7% of the population) becoming infected by the age of 10. Linkage analysis was carried out on the 15 most informative families, corresponding to 205 genotyped subjects. A single region on chromosome 3p22 was significantly linked to HHV-8 infection (LOD score=3.83, P=2.0 × 10(-5)). This study provides the first evidence that HHV-8 infection in children in endemic areas has a strong genetic basis involving at least one recessive major locus on chromosome 3p22.


Acridine orange staining of smears for demonstration of Mycobacterium tuberculosis.
Acridine orange staining for the detection of mycobacteria was compared with staining by auramine O and with mycobacterial culture in a series of 1071 clinical specimens. A total of 78 (7%) specimens were positive by staining. No false positive or negative findings were recorded by the acridine orange method. The two fluorochromes proved equal in their ability to detect mycobacteria in specimens from culture positive cases of tuberculosis. In the rapid bacteriological diagnosis of tuberculosis, acridine orange offers a good alternative to auramine O which is considered carcinogenic.


Time-related decreases in mu and delta opioid receptors in the superficial dorsal horn of the rat spinal cord following a large unilateral dorsal rhizotomy.
The aim of the present study was to measure the time-related modifications of mu and delta opioid binding sites in the superficial layers of the dorsal horn of the rat spinal cord after a C4-T2 unilateral dorsal rhizotomy. Using specific ligands, namely [3H]DAMGO for mu sites and [3H]DTLET for delta sites, and a quantitative autoradiographic analysis, we have observed: (a) a decrease in binding on the ipsilateral side to the lesion as early as the first day postrhizotomy, the maximal loss being attained at 8 days postlesion, (b) after 8 days postlesion, the residual binding remains stable over the period of analysis (90 days), (c) the loss of mu receptors (71-74%) is significantly more pronounced than the loss of delta receptors (57-62%) and (d) affinities of postsynaptic mu and delta receptors are similar to those of the total receptor population in the superficial layers of the dorsal horn. Comparison of these results with the degeneration of primary afferent fibers reported in literature favors the localization of the majority of mu and delta opioid binding sites on fine diameter primary afferent fibers.


High Performance Computing Framework for Tera-Scale Database Search of Mass Spectrometry Data.
Database peptide search algorithms deduce peptides from mass spectrometry (MS) data. There has been substantial effort in improving their computational efficiency to achieve larger and more complex systems biology studies. However, modern serial and high-performance computing (HPC) algorithms exhibit sub-optimal performance mainly due to their ineffective parallel designs (low resource utilization), and high overhead costs. We present an HPC framework, called HiCOPS, for efficient acceleration of the database peptide search algorithms on distributed-memory supercomputers. HiCOPS provides, on average, more than 10-fold improvement in speed, and superior parallel performance over several existing HPC database search software. We also formulate a mathematical model for performance analysis and optimization, and report near-optimal results for several key metrics including strong-scale efficiency, hardware utilization, load-balance, inter-process communication and I/O overheads. The core parallel design, techniques, and optimizations presented in HiCOPS are search-algorithm independent and can be extended to efficiently accelerate the existing and future algorithms and software.


Public health indicators for the EU: the joint action for ECHIM (European Community Health Indicators & Monitoring).
Public health policies aim to improve and maintain the health of citizens. Relevant data and indicators are needed for a health policy that is based on factual information. After 14 years of work (1998-2012), the multi-phase action on European Community Health Indicators (ECHI) has created a health monitoring and reporting system. It has generated EU added value by defining the ECHI shortlist with 88 common and comparable key health indicators for Europe. In the 2009-2012 Joint Action for ECHIM project the ECHI shortlist was updated through consultation with Member State representatives. Guidelines for implementation of the ECHI Indicators at national level were developed and a pilot data collection was carried out. 67 of the ECHI Indicators are already part of regular international data collections and thus available for a majority of Member States, 14 are close to ready and 13 still need development work. By mid-2012 half of the countries have incorporated ECHI indicators in their national health information systems and the process is ongoing in the majority of the countries. Twenty-five countries were able to provide data in a Pilot Data Collection for 20 ECHI Indicators that were not yet (fully) available in the international databases. The EU needs a permanent health monitoring and reporting system. The Joint Action for ECHIM has set an example for the implementation of a system that can develop and maintain the ECHI indicators,, and promote and encourage the use of ECHI in health reporting and health policy making. The aim for sustainable public health monitoring is also supported by a Eurostat regulation on public health statistics requiring that health statistics shall be provided according to the ECHI methodology. Further efforts at DG SANCO and Eurostat are needed towards a permanent health monitoring system.


Association of white matter microstructure and extracellular free-water with cognitive performance in the early course of schizophrenia.
Schizophrenia (SZ) is proposed as a disorder of dysconnectivity underlying cognitive impairments and clinical manifestations. Although previous studies have shown extracellular changes in white matter of first-episode SZ, little is known about the transition period towards chronicity and its association with cognition. Free-water (FW) imaging was applied to 79 early course SZ participants and 29 controls to detect white matter axonal and extracellular differences during this phase of illness. Diffusion-weighted images were collected from two sites, harmonized, and processed using a pipeline separately modeling water diffusion in tissue (FAt) and extracellular space (FW). Tract-Based Spatial Statistics was performed using the ENIGMA-DTI protocols. SZ showed FAt reductions in the posterior thalamic radiation (PTR) and FW elevations in the cingulum compared to controls, suggesting FAt and FW changes in the early course of SZ. In SZ, greater FAt of the fornix & stria terminalis (FXST) was positively associated with Theory of Mind performance; average whole-brain FAt, FAt of the FXST and the PTR were positively associated with greater working memory performance; average whole-brain FAt was positively associated with visual learning. Further studies are necessary to better understand the neurobiological mechanisms of SZ for developing intervention strategies to preserve brain structure and function.


NLRP7 participates in the human subcortical maternal complex and its variants cause female infertility characterized by early embryo arrest.
Successful human reproduction requires normal oocyte maturation, fertilization, and early embryo development. Early embryo arrest is a common phenomenon leading to female infertility, but the genetic basis is largely unknown. NLR family pyrin domain-containing 7 (NLRP7) is a member of the NLRP subfamily. Previous studies have shown that variants of NLRP7 are one of the crucial causes of female recurrent hydatidiform mole, but whether NLRP7 variants can directly affect early embryo development is unclear. We performed whole-exome sequencing in patients who experienced early embryo arrest, and five heterozygous variants (c.251G > A, c.1258G > A, c.1441G > A, c. 2227G > A, c.2323C > T) of NLRP7 were identified in affected individuals. Plasmids of NLRP7 and subcortical maternal complex components were overexpressed in 293 T cells, and Co-IP experiments showed that NLRP7 interacted with NLRP5, TLE6, PADI6, NLRP2, KHDC3L, OOEP, and ZBED3. Injecting complementary RNAs in mouse oocytes and early embryos showed that NLRP7 variants influenced the oocyte quality and some of the variants significantly affected early embryo development. These findings contribute to our understanding of the role of NLRP7 in human early embryo development and provide a new genetic marker for clinical early embryo arrest patients. KEY MESSAGES: Five heterozygous variants of NLRP7 (c.1441G > A; 2227G > A; c.251G > A; c.1258G > A; c.2323C > T) were identified in five infertile patients who experienced early embryo arrest. NLRP7 is a component of human subcortical maternal complex. NLRP7 variants lead to poor quality of oocytes and early embryo development arrest. This study provides a new genetic marker for clinical early embryo arrest patients.


Spectroscopic evidence for the generation of singlet oxygen in the reduced nicotinamide adenine dinucleotide phosphate-dependent microsomal lipid peroxidation system.
In the presence of Fe3+-ADP and NADPH, washed rat liver microsomes at 37 degrees and at pH 7.5 emitted ultraweak light with the peroxidative cleavage of endogenous lipid. The main light-emitting species of this system was analyzed spectroscopically in the visible region and estimated to be singlet oxygen. The addition of superoxide dismutase or catalase did not inhibit the light emission significantly. It is unlikely therefore that superoxide anion and OH are involved in the generation of singlet oxygen in the present system.


Synthesis and cytotoxic activity of lipophilic sulphonamide derivatives of the benzo[b]thiophene 1,1-dioxide.
In the search of new compounds with antineoplastic activity, we have analysed the effect of several structural modifications on the nucleus 6-benzo[b]thiophenesulphonamide 1,1-dioxide on its cytotoxic activity on tumour cells. Lipophilic substituents on the sulphonamide group significantly increased the cytotoxic activity measured using a panel of human tumour cell lines. Only slight variations on cytotoxicity were obtained when the sulphonamide group occupied the position 5 of the system. The most active compound was the N-4-methoxyphenyl derivative 15, which showed GI(50) values of 1-9 nM against HT-29, CCRF-CEM, K-562 and MEL-AC cells and of 200 nM against HTB-54 cells. Free access to the 3-position of the heterocyclic system seems to be required to obtain cytotoxic derivatives. Derivative 15 was also active at the same level of commercial Doxorubicine against cultured normal human lung fibroblasts.


Levels of Intervention: How Are They Used in Quebec Hospitals?
In order to promote better practices and communication around end-of-life decision-making, several Canadian hospitals in the province of Quebec have developed a tool called "Levels of Intervention" (LOI). No work to date has been published demonstrating improvement since these forms were implemented. The purpose of the present study was to obtain information about the use of LOI forms across Quebec hospitals and to identify gaps in practice as well as areas for improvement. A retrospective study was undertaken of 299 charts of patients who had died in three Quebec hospitals with a LOI ordered. Results were analysed through a principlism ethical framework. High compliance with the level of intervention ordered at the time of death was recorded, as well as high involvement of patient and/or family, demonstrating the efficiency of LOI in promoting respect for autonomy. Other results show delays in end-of-life care discussions in the course of the hospitalization. Only a small proportion of patients who died had a palliative care consultation, which may reflect equity issues in access to care. This study highlights the importance of the LOI in Quebec and the role it is playing in respect for end-of-life preferences as well as in the involvement of patients and families in the decision-making process. Training specific to end-of-life decision-making conversations would help support the LOI form's use, as would developing provincial or national guidelines on the use of LOI to standardize organizational policies and practice around end-of-life care.


P143 proteins from heterologous nucleopolyhedroviruses induce apoptosis in BM-N cells derived from the silkworm Bombyx mori.
We previously demonstrated that ribosomal RNA (rRNA) of Bombyx mori BM-N cells is rapidly degraded upon infection with heterologous nucleopolyhedroviruses (NPVs), including Autographa californica multiple NPV (AcMNPV), Hyphantria cunea MNPV, Spodoptera exigua MNPV and S. litura MNPV, and that this response is triggered by viral P143 proteins. The transient expression of P143 proteins from heterologous NPVs was also shown to induce apoptosis and caspase-3-like protease activation in BM-N cells. In the present study, we conducted a transient expression assay using BM-N cells expressing mutant AcMNPV P143 (Ac-P143) proteins and demonstrated that five amino acid residues cooperatively participate in Ac-P143 protein-triggered apoptosis of BM-N cells. Notably, these five residues were previously shown to be required for triggering rRNA degradation in BM-N cells. As rRNA degradation in BM-N cells does not result from apoptosis, the present results suggest that Ac-P143-triggered rRNA degradation is the upstream signal for apoptosis induction in BM-N cells. We further showed that P143 protein-triggered apoptosis does not occur in S. frugiperda Sf9 or Lymantria dispar Ld652Y cells, indicating that apoptosis induction by heterologous P143 proteins is a BM-N cell-specific response. In addition, the observed induction of apoptosis in BM-N cells was found to be mediated by activation of the initiator caspase Bm-Dronc. Taken together, these results suggest that BM-N cells evolved a unique antiviral system that recognizes heterologous NPV P143 proteins to induce rRNA degradation and caspase-dependent apoptosis.


Treatment sequencing using the dual amylin and calcitonin receptor agonist KBP-336 and semaglutide results in durable weight loss.
Long-acting dual amylin and calcitonin receptor agonists (DACRAs) hold great promise as potential treatments for obesity and its associated comorbidities. These agents have demonstrated beneficial effects on body weight, glucose control, and insulin action mirroring the effects observed with glucagon-like peptide-1 (GLP-1) agonist treatment. Strategies aimed at enhancing and prolonging treatment efficacy include treatment sequencing and combination therapy. Here, we sought to investigate the impact of switching between or combining treatment with the DACRA KBP-336 and the GLP-1 analog semaglutide in fed rats with obesity induced by a high-fat diet (HFD). Two studies were performed in which HFD-induced obese Sprague Dawley rats were switched between treatment with KBP-336 (4.5 nmol/kg, Q3D) and semaglutide (50 nmol/kg, Q3D) or a combination of the two. Treatment efficacy on weight loss and food intake was evaluated, and glucose tolerance was assessed by oral glucose tolerance tests. KBP-336 and semaglutide monotherapy resulted in a similar reduction in body weight and food intake. Treatment sequencing resulted in continuous weight loss and all monotherapies resulted in similar weight loss independent of the treatment regimen (P < 0.001 compared to vehicle). The combination of KBP-336 and semaglutide significantly improved the weight loss compared to either monotherapy alone (P < 0.001), which was evident in the adiposity at the study end. All treatments improved glucose tolerance, with the KBP-effect on insulin sensitivity as the dominant response. These findings highlight KBP-336 as a promising anti-obesity therapy both alone, in treatment sequencing, and in combination with semaglutide or other incretin-based therapies.


The EutQ and EutP proteins are novel acetate kinases involved in ethanolamine catabolism: physiological implications for the function of the ethanolamine metabolosome in Salmonella enterica.
Salmonella enterica catabolizes ethanolamine inside a compartment known as the metabolosome. The ethanolamine utilization (eut) operon of this bacterium encodes all functions needed for the assembly and function of this structure. To date, the roles of EutQ and EutP were not known. Herein we show that both proteins have acetate kinase activity and that EutQ is required during anoxic growth of S. enterica on ethanolamine and tetrathionate. EutP and EutQ-dependent ATP synthesis occurred when enzymes were incubated with ADP, Mg(II) ions and acetyl-phosphate. EutQ and EutP also synthesized acetyl-phosphate from ATP and acetate. Although EutP had acetate kinase activity, ΔeutP strains lacked discernible phenotypes under the conditions where ΔeutQ strains displayed clear phenotypes. The kinetic parameters indicate that EutP is a faster enzyme than EutQ. Our evidence supports the conclusion that EutQ and EutP represent novel classes of acetate kinases. We propose that EutQ is necessary to drive flux through the pathway under physiological conditions, preventing a buildup of acetaldehyde. We also suggest that ATP generated by these enzymes may be used as a substrate for EutT, the ATP-dependent corrinoid adenosyltransferase and for the EutA ethanolamine ammonia-lyase reactivase.


A Single-Transaminase-Catalyzed Biocatalytic Cascade for Efficient Asymmetric Synthesis of l-Phosphinothricin.
A single-transaminase-catalyzed biocatalytic cascade was developed by employing the desired biocatalyst, ATA-117-Rd11, that showed high activity toward 2-oxo-4-[(hydroxy)(methyl)phosphinoyl] butyric acid (PPO) and α-ketoglutarate, and low activity against pyruvate. The cascade successfully promotes a highly asymmetric amination reaction for the synthesis of l-phosphinothricin (l-PPT) with high conversion (>95 %) and>99 % ee. In a scale-up experiment, using 10 kg pre-frozen E. coli cells harboring ATA-117-Rd11 as catalyst, 80 kg PPO was converted to ≈70 kg l-PPT after 24 hours with a high ee value (>99 %).


Impact of different interventions on preventing suicide and suicide attempt among children and adolescents in the United States: a microsimulation model study.
Despite considerable investment in suicide prevention since 2001, there is limited evidence for the effect of suicide prevention interventions among children and adolescents. This study aimed to estimate the potential population impact of different interventions in preventing suicide-related behaviors in children and adolescents. A microsimulation model study used data from national surveys and clinical trials to emulate the dynamic processes of developing depression and care-seeking behaviors among a US sample of children and adolescents. The simulation model examined the effect of four hypothetical suicide prevention interventions on preventing suicide and suicide attempt in children and adolescents as follows: (1) reduce untreated depression by 20, 50, and 80% through depression screening; (2) increase the proportion of acute-phase treatment completion to 90% (i.e., reduce treatment attrition); (3) suicide screening and treatment among the depressed individuals; and (4) suicide screening and treatment to 20, 50, and 80% of individuals in medical care settings. The model without any intervention simulated was the baseline. We estimated the difference in the suicide rate and risk of suicide attempts in children and adolescents between baseline and different interventions. No significant reduction in the suicide rate was observed for any of the interventions. A significant decrease in the risk of suicide attempt was observed for reducing untreated depression by 80%, and for suicide screening to individuals in medical settings as follows: 20% screened: -0.68% (95% credible interval (CI): -1.05%, -0.56%), 50% screened: -1.47% (95% CI: -2.00%, -1.34%), and 80% screened: -2.14% (95% CI: -2.48%, -2.08%). Combined with 90% completion of acute-phase treatment, the risk of suicide attempt changed by -0.33% (95% CI: -0.92%, 0.04%), -0.56% (95% CI: -1.06%, -0.17%), and -0.78% (95% CI: -1.29%, -0.40%) for reducing untreated depression by 20, 50, and 80%, respectively. Combined with suicide screening and treatment among the depressed, the risk of suicide attempt changed by -0.27% (95% CI: -0.dd%, -0.16%), -0.66% (95% CI: -0.90%, -0.46%), and -0.90% (95% CI: -1.10%, -0.69%) for reducing untreated depression by 20, 50, and 80%, respectively. Reducing undertreatment (the untreated and dropout) of depression and suicide screening and treatment in medical care settings may be effective in preventing suicide-related behaviors in children and adolescents.


Polymorphonuclear leukocytes in brain parenchyma after injury and their interaction with purified astrocytes in culture.
At a time after brain injury when removal of debris and secondary cell death were prevalent, many polymorphonuclear neutrophils were observed in injured tissue. Because neural damage could be mediated by activated neutrophils, we tested in vitro the effect of these leukocytes and other blood components on central nervous system cells. At concentrations similar to those present in blood, polymorphonuclear leukocytes resulted in astrocyte detachment from the substrate and aggregation. These neutrophil concentrations affected both epithelioid and stellate HNK1/A2B5-negative (type 1) astrocytes but not hippocampal neurons. Substrate detachment was partially prevented by corticosterone, but not by protease inhibitors or free-radical scavengers. Co-cultures of purified cortical astrocytes with neutrophils (1/20 cell ratio) contained at the beginning of the experiment approximately 93-98% astrocytes with type 1 markers. After 6-8 days co-culture, many stellate cells insensitive to neutrophils seemed to migrate out of the aggregates. About 70% of these resistant cells had immunological markers typical of type 2 astrocytes. The possible relevance of these findings to reactive astrogliosis and secondary neuronal death is discussed.


Optic disk and visual field correlations in primary open-angle and low-tension glaucoma.
If the amount of visual field loss is less than expected from the amount of optic disk cupping in low-tension glaucoma compared with primary open-angle glaucoma, it might imply a difference between the two conditions in the type of optic nerve lesion produced. To test this hypothesis, three observers independently examined, in a masked fashion, optic disk stereoscopic photographs of 127 eyes with primary open-angle glaucoma and 71 eyes with low-tension glaucoma. For each stereoscopic photograph the observer predicted whether the visual field loss would be mild, moderate, or severe. The visual field were then classified, according to the number of sectors defective on the Goldmann perimeter chart, as having mild (1 to 15 sectors), moderate (16 to 30 sectors), or severe (more than 30 sectors) visual field loss. For no observer did the frequency of underpredictions or overpredictions in the two conditions differ significantly. The results of this study, thus, did not support the theory that the optic disk damage in primary open-angle glaucoma differs from that in low-tension glaucoma.


Distinct heterogeneity of IgG immune response in cerebrospinal fluid (CSF) detected by isoelectric focusing (IEF) with extended immunofixation.
An optimized automated IEF procedure in polyacrylamide micro gels and immunofixation with 10 monospecific antibodies against some fragments of the IgG molecule and against the whole IgG isotypes IgG(1), IgG(2), IgG(3), IgG(4), respectively, detected oligoclonal bands (OBs) within acid, neutral, and alkaline ranges of the gels. Accuracy and reliability of the OB assay for detection of an intrathecal IgG synthesis proved to be higher with immunofixation than with silver staining of bands precipitated by trichloroacetic acid. CSF OBs were specified as lambda or kappa IgG subfractions, respectively, duplex IgGs precipitated with anti-lambda, anti-kappa, and anti-Fc antibodies at the same pI. Most of OBs were classified as belonging to either IgG(1), IgG(2), IgG(3), or IgG(4) isotypes in CSF. The IEF procedure additionally allowed the discrimination of both free light chains and possible "free" heavy gamma chain fragments in CSF, when immunofixation was done with monospecific antibodies against both light chains and gamma chain fragments (e.g. anti-F(ab')(2), anti-Fd, anti-Fc, anti-C(H)2). The results pointed out a distinct heterogeneity of the IgG immune response in human CSF. The IEF procedure with extended immunofixation is recommended for completion of the basic laboratory procedures used in neuroimmunology in order to discriminate inflammatory processes in human CNS.


Medicare program; standards for e-prescribing under Medicare Part D and identification of backward compatible version of adopted standard for e-prescribing and the Medicare prescription drug program (version 8.1). Final rule.
This final rule adopts uniform standards for medication history, formulary and benefits, and fill status notification (RxFill) for the Medicare Part D electronic prescribing (e-prescribing) drug program as required by section 1860D-4(e)(4)(D) of the Social Security Act (the Act). In addition, we are adopting the National Provider Identifier (NPI) as a standard for identifying health care providers in e-prescribing transactions. It also finalizes the June 23, 2006 interim final rule with comment period that identified the National Council for Prescription Drug Programs (NCPDP) Prescriber/Pharmacist Interface SCRIPT standard, Implementation Guide, Version 8.1 ("NCPDP SCRIPT 8.1") as a backward compatible update of the NCPDP SCRIPT 5.0 ("NCPDP SCRIPT 5.0"), until April 1, 2009. This final rule also retires NCPDP SCRIPT 5.0 and adopts the newer version, NCPDP SCRIPT 8.1, as the adopted standard. Finally, except as otherwise set forth herein, we are implementing our compliance date of 1 year after the publication of these final uniform standards. This is the second set in a continuing process of issuing e-prescribing final standards for the Medicare Part D program,


Combined small RNA and gene expression analysis revealed roles of miRNAs in maize response to rice black-streaked dwarf virus infection.
Maize rough dwarf disease, caused by rice black-streaked dwarf virus (RBSDV), is a devastating disease in maize (Zea mays L.). MicroRNAs (miRNAs) are known to play critical roles in regulation of plant growth, development, and adaptation to abiotic and biotic stresses. To elucidate the roles of miRNAs in the regulation of maize in response to RBSDV, we employed high-throughput sequencing technology to analyze the miRNAome and transcriptome following RBSDV infection. A total of 76 known miRNAs, 226 potential novel miRNAs and 351 target genes were identified. Our dataset showed that the expression patterns of 81 miRNAs changed dramatically in response to RBSDV infection. Transcriptome analysis showed that 453 genes were differentially expressed after RBSDV infection. GO, COG and KEGG analysis results demonstrated that genes involved with photosynthesis and metabolism were significantly enriched. In addition, twelve miRNA-mRNA interaction pairs were identified, and six of them were likely to play significant roles in maize response to RBSDV. This study provided valuable information for understanding the molecular mechanism of maize disease resistance, and could be useful in method development to protect maize against RBSDV.


[Etiologies of papilledema in Cameroonian hospitals].
papilledema refers to the swelling of the head of the optic nerve, a major sign of many local, locoregional or systemic pathologies that may involve patients' visual or vital prognosis. This condition represents a diagnostic and therapeutic emergency frequently encountered in our facilities. Therefore, it is deemed and opportune to identify the most common etiologies observed in Cameroonian hospitals. we conducted a documentary and descriptive study of patients with papilledema presenting to the Ophthalmology Department of the Hospital of Instruction, Application and Reference of the Armed Forces of Yaoundé from 1st October 2013 to 31st December 2016. The variables under investigation included epidemiological data (age, sex), clinical data (functional signs, visual acuity, appearance of the papilla and associated signs according to the Hoyt and Beesten classification), complementary examinations performed (fluorescein angiography, visual field, biology, radiography, CT scan) and the etiological diagnosis. Epi-info 3.5.3 software was used for statistical analysis and the Chi-square test was performed at a 5% significance level (p < 5%). during the study period, papilledema was found in 26 out of 5023 patients, reflecting a rate of 0.5%. The average age of patients was 32.7± 10.9 years, ranging from 7 to 79 years, for 13 women and 13 men. Papilledema was bilateral in 15 (57.7%) patients and unilateral in 11 (42.3%), or 41 affected eyes. The etiologies were 11 (42,3%) inflammatory optic neuropathies, 5 (19,2%) arterial hypertension, 4 (15,4%) central retinal vein occlusions, 3 (11,5%) eye contusions, 2 (7,7%) hydrocephalus and 1 case (3,9%) of cerebral malaria. inflammatory and vascular optic neuropathies were the most common etiologies of papilledema in our facilities.


A comprehensive three-dimensional assay to assess neutrophil defense against bacteria.
Neutrophil antibacterial capacity is measured in animal models and in vitro as an important indicator of neutrophil function. To be able to extrapolate their conclusions, in vitro experiments should mimic the in vivo situation. In vivo, antibacterial capacity depends on multiple steps of bacterial sensing, priming, chemotaxis, phagocytosis and intracellular killing. Therefore, we developed a simply executed assay that involves multiple steps in one assay. The neutrophils were incorporated into a three-dimensional matrix of fibrin fibers, in which they could freely migrate. The fibrin matrix provided a more physiological representation of tissue structure than a shaken suspension and extended ex vivo survival of neutrophils. Staphylococci endogenously producing GFP (Green Fluorescent Protein) provided a real-time quantification of the bacterial load without the need for lysing the fibrin matrix or counting of colony forming units on agar plates. The delay in bacterial outgrowth serves as a measure for the relative antibacterial capacity of the neutrophils. Additionally, neutrophil capacity could easily be measured high-throughput in a 96-wells format. In this new assay we study neutrophil behavior in a physiologically relevant setting and explore many functions of the neutrophil in a single test. The functional capacity of neutrophils from different in vitro treatments or different donors can directly be compared.


Structure and function of a benthic invertebrate stream community as influenced by beaver (Castor canadensis).
Beaver (Castor canadensis) affect the benthic invertebrate community of small woodland streams in Quebec through habitat modifications. Their activities influence community structure through the replacement of lotic taxa by lentic forms and community function by increasing the absolute importance of collectors and predators while decreasing the relative importance of shredders and scrapers in impounded sites. At our study site during the 1983 ice-free season, standing stocks of coarse particulate organic matter (>1 mm) were 2-5 times greater (P<0.05) in impounded sites than riffle sites in spring and summer. Fine (212 μm-1 mm) and very fine (0.5 μm-212 μm) particulate organic matter were 3-10 times greater (P<0.05) in impounded sites in all seasons. Chlorophyll a standing stocks did not differ statistically among sites. Total density and biomass of invertebrates in impoundments were 2-5 times greater (P<0.05) than riffle sites in spring and summer, but statistically similar in autumn. Generic diversity (H') was greater (P<0.05) in unaltered sites in autumn. Non-impounded sites were dominated by Simuliidae, Tanytarsini chironomids, scraping mayflies and net spinning caddisflies while impounded sites were characterized by Tanypodinae and Chironomini chironomids, predacious odonates, Tubificidae, and filtering pelycopods. Our results suggest that current paradigms applied to lotic ecosystems need to be reevaluated to incorporate the influence of beaver upon invertebrate communities.


Long-term event-free survivors after high-dose therapy and autologous stem-cell transplantation for low-grade follicular lymphoma.
Although follicular lymphoma (FL) is generally responsive to conventional-dose chemotherapy, improved survival in patients with this disease has been difficult to demonstrate. High-dose chemo/radiotherapy followed by autologous stem-cell transplantation (ASCT) can improve response rates, although its effects on survival remain controversial. Between 1990 and 2003, we transplanted 49 patients with low-grade FL at our institution. Twenty-two patients (45%) had undergone histologic transformation at the time of ASCT. In all, 44 patients (90%) had relapsed disease and five patients (10%) were resistant to chemotherapy at the time of transplantation. After ASCT, 30 patients (61%) were in complete remission (CR). The median overall survival (OS) has not been reached, while the median event-free survival (EFS) is 2.4 years. At a median follow-up of 5.5 years (longest 12.4 years), a plateau has been reached with 56% of patients remaining alive, and 35% event-free. ASCT was well tolerated except for two (4%) treatment-related deaths. In multivariable analysis, CR after ASCT and age less than 60 years are the best predictors of EFS and OS. ASCT is thus a safe therapeutic approach in FL, resulting in long-term EFS and OS for some patients, even with transformed disease.


Outcome of interspecific competition depends on genotype of conspecific neighbours.
The outcome of interspecific competition is affected by numerous abiotic and biotic factors. However, the effects of genetic relatedness of conspecific neighbours have not been elucidated. Here, we demonstrate that the genotype of a conspecific neighbour determines the outcome of interspecific competition. We investigated effects of genotype of neighbour sibling or non-sibling Plantago asiatica plants on competition with Trifolium repens plants. Pairs of P. asiatica, sibling or non-sibling, were grown in competition with T. repens. Sibling P. asiatica plants, but not non-sibling plants, increased leaf placement towards competitor T. repens plants, and that placement was correlated negatively with the biomass of T. repens and positively with seed production by P. asiatica. Seed production by P. asiatica plants competing with T. repens was greater in individuals paired with siblings than in those paired with non-siblings. Thus, our results show that the genotype of conspecific neighbours can alter the outcome of interspecific competition.


Altered thioredoxin subcellular localization and redox status in MCF-7 cells following 1,25-dihydroxyvitamin D3 treatment.
1,25-Dihydroxyvitamin D(3) (1,25D) induces apoptosis in MCF-7 cells via the intrinsic pathway involving bax translocation to mitochondria, cytochrome c release and reactive oxygen species (ROS) generation. Vitamin D up-regulated protein 1 (VDUP1), an apoptotic regulatory gene induced by 1,25D in HL-60 cells, is a negative regulator of thioredoxin (Trx1), a redox protein which neutralizes ROS and protects against oxidative stress induced apoptosis. Due to the involvement of oxidative stress in 1,25D mediated apoptosis, we analyzed whether VDUP1 or Trx1 are altered by 1,25D in MCF-7 cells. In contrast to HL-60 cells, VDUP1 mRNA was not up-regulated by 1,25D in MCF-7 cells, indicating that transcriptional up-regulation of this gene is not required for 1,25D mediated apoptosis. 1,25D did not affect the expression or activity of Trx1 in MCF-7 cells, however, Trx1 activity was higher in MCF-7 cells selected for resistance to 1,25D mediated apoptosis. In untreated MCF-7 cells, Trx1 was present only in the cytosol, and the majority was in the oxidized state. In 1,25D treated MCF-7 cells, Trx1 was present in both cytosol and nucleus, and the nuclear Trx1 pool was in the reduced state. Nuclear localization of Trx1 in 1,25D treated MCF-7 cells was confirmed by immunofluorescent microscopy. Although redox status is known to alter the ability of Trx1 to bind apoptosis signal regulating kinase 1 (ASK1), no changes in ASK1 transcript or protein levels were observed in 1,25D treated MCF-7 cells. Collectively, these studies indicate that although VDUP1 and ASK1 are not altered by 1,25D, changes in redox status and sub-cellular distribution of Trx1 occurs during 1,25D mediated apoptosis of MCF-7 cells.


Properties of docosahexaenoic-acid-containing lipids and their influence on the function of rhodopsin.
The importance of highly polyunsaturated fatty acids in health and development has been convincingly demonstrated by many studies over the past several decades. The mechanisms by which polyunsaturated lipid species might influence biological function at the molecular level are now attracting considerable attention. The G-protein-coupled receptor rhodopsin and docosahexaenoic acid, the dominant fatty acid in the retinal membrane, provide the best-studied example of protein function being influenced by lipid environment.


Nutritional supplementation in monochorionic diamniotic twin pregnancies: impact on twin-twin transfusion syndrome.
Twin-to-twin transfusion syndrome (TTTS) has been related to unbalanced unidirectional arteriovenous anastomoses in the placenta of monochorionic diamniotic (DiMo) twin gestations. As maternal malnutrition accounting for hypoproteinemia and anemia has been detected in severe cases of TTTS, the purpose of this study was to evaluate the impact of early diet supplementation on TTTS. Fifty-one DiMo twin pregnancies were given commercially available oral nutritional diet supplements and then compared in a retrospective cohort study to 52 twin gestations with the same chorionicity but not subjected to nutritional supplementation. Diet supplementation was associated with lower overall incidence of TTTS (20/52 versus 8/51, P = 0.02) and with lower prevalence of TTTS at delivery (18/52 versus 6/51, P = 0.012) when compared with no supplementation. Nutritional intervention also significantly prolonged the time between the diagnosis of TTTS and delivery (9.4 +/- 3.7 weeks versus 4.6 +/- 6.5 weeks; P = 0.014). The earlier nutritional regimen was introduced, the lesser chance of detecting TTTS ( P = 0.001). Although not statistically significant, dietary intervention was also associated with lower Quintero stage, fewer invasive treatments, and lower twin birth weight discordance. Diet supplementation appears to counter maternal metabolic abnormalities in DiMo twin pregnancies and improve perinatal outcomes in TTTS when combined with the standard therapeutic options.