[Multidisciplinary approach of hepatic metastasis of colorectal origin].
Surgery remains the only potentially curative treatment for liver metastases. After margin free resection, approximately 30% of the patients present long-term survival. Due to the metastases number and/or volume, only 10 to 15% of the patients are candidates for curative-intent surgery. Thus, the objectives of the diagnostic and therapeutic management are to select adequately the patients for surgery and to improve resection rate by the use of neoadjuvants methods. Positron emission tomography could improve the preoperative detection of hepatic and extrahepatic metastases leading to the exclusion of some patients from useless surgical exploration. For patients with initially resectable tumors, no benefit has been demonstrated for adjuvant chemotherapy. For non resectable metastases, two neoadjuvants methods should be evaluated, chemotherapy to reduce tumor volume and portal vein embolization of the tumor side to improve the hepatic functional reserve and allow larger resection. For non-accessible lesion, selective tumor destruction using radiofrequency offers promising perspectives. In conclusion, the multiplication of the diagnostic and therapeutic methods certainly improve the global management of patients with liver metastases but also makes more difficult the individual choice for the best treatment. For this reason, a multimodal approach is absolutely mandatory.


Combined single-session cardiovascular magnetic resonance: stress perfusion and three-dimensional pulmonary vein angiography for stratification of atrial fibrillation patients with chest pain syndromes prior to catheter ablation.
To determine the clinical utility of a combined single-session cardiovascular magnetic resonance (CMR) imaging protocol integrating adenosine stress perfusion and three-dimensional pulmonary vein angiography for stratification of atrial fibrillation (AF) patients referred for pulmonary vein isolation (PVI) and complaining about chest pain syndromes. The preprocedural CMR examination (adenosine stress perfusion, late gadolinium enhancement, and three-dimensional pulmonary vein angiography) was performed in 357 consecutive AF patients with chest pain syndromes referred for PVI. Stress perfusion results were used for stratification: ischaemia positive patients underwent invasive coronary angiography, ischaemia negative patients underwent PVI, and follow-up/outcome data were collected (combined primary endpoint of cardiac death/non-fatal myocardial infarction). The integrated CMR protocol had a high success rate (356/357, 99.7%), a short total examination duration (<30 min in all patients), and delivered high-quality three-dimensional pulmonary vein angiography in all patients undergoing PVI (324/324, 100%). Variants of pulmonary vein anatomy were identified in 33% of all patients (117/357). Stress positivity (28/356, 8%) had a high positive predictive value for identification of obstructive coronary artery disease (86%), while stress negativity carried a low short-term event rate following PVI (cumulative 1-year event-free survival rate, 99.6%). Combined single-session CMR as a routine diagnostic workup for AF patients with chest pain syndromes prior to PVI proved to represent a time-efficient and effective stratification tool.


Computational repositioning and preclinical validation of pentamidine for renal cell cancer.
Although early stages of clear cell renal cell carcinoma (ccRCC) are curable, survival outcome for metastatic ccRCC remains poor. We previously established a highly accurate signature of differentially expressed genes that distinguish ccRCC from normal kidney. The purpose of this study was to apply a new individualized bioinformatics analysis (IBA) strategy to these transcriptome data in conjunction with Gene Set Enrichment Analysis of the Connectivity Map (C-MAP) database to identify and reposition FDA-approved drugs for anticancer therapy. Here, we demonstrate that one of the drugs predicted to revert the RCC gene signature toward normal kidney, pentamidine, is effective against RCC cells in culture and in a RCC xenograft model. ccRCC-specific gene expression signatures of individual patients were used to query the C-MAP software. Eight drugs with negative correlation and P-value <0.05 were analyzed for efficacy against RCC in vitro and in vivo. Our data demonstrate consistency across most patients with ccRCC for the set of high-scoring drugs. Most of the selected high-scoring drugs potently induce apoptosis in RCC cells. Several drugs also demonstrate selectivity for Von Hippel-Lindau negative RCC cells. Most importantly, at least one of these drugs, pentamidine, slows tumor growth in the 786-O human ccRCC xenograft mouse model. Our findings suggest that pentamidine might be a new therapeutic agent to be combined with current standard-of-care regimens for patients with metastatic ccRCC and support our notion that IBA combined with C-MAP analysis enables repurposing of FDA-approved drugs for potential anti-RCC therapy.


Multiple sclerosis-a quiet revolution.
Multiple sclerosis (MS) has been thought to be a complex and indecipherable disease, and poorly understood with regards to aetiology. Here, we suggest an emphatically positive view of progress over several decades in the understanding and treatment of MS, particularly focusing on advances made within the past 20 years. As with virtually all complex disorders, MS is caused by the interaction of genetic and environmental factors. In recent years, formidable biochemical, bioinformatic, epidemiological and neuroimaging tools have been brought to bear on research into the causes of MS. While susceptibility to the disease is now relatively well accounted for, disease course is not and remains a salient challenge. In the therapeutic realm, numerous agents have become available, reflecting the fact that the disease can be attacked successfully at many levels and using varied strategies. Tailoring therapies to individuals, risk mitigation and selection of first-line as compared with second-line medications remain to be completed. In our view, the MS landscape has been comprehensively and irreversibly transformed by this progress. Here we focus on MS therapeutics-the most meaningful outcome of research efforts.


Requirements for infrastructure and essential activities of infection control and epidemiology in hospitals: A consensus panel report. Society for Healthcare Epidemiology of America.
The scientific basis for claims of efficacy of nosocomial infection surveillance and control programs was established by the Study on the Efficacy of Nosocomial Infection Control project. Subsequent analyses have demonstrated nosocomial infection prevention and control programs to be not only clinically effective but also cost-effective. Although governmental and professional organizations have developed a wide variety of useful recommendations and guidelines for infection control, and apart from general guidance provided by the Joint Commission on Accreditation of Healthcare Organizations, there are surprisingly few recommendations on infrastructure and essential activities for infection control and epidemiology programs. In April 1996, the Society for Healthcare Epidemiology of America established a consensus panel to develop recommendations for optimal infrastructure and essential activities of infection control and epidemiology programs in hospitals. The following report represents the consensus panel's best assessment of needs for a healthy and effective hospital-based infection control and epidemiology program. The recommendations fall into eight categories: managing critical data and information; setting and recommending policies and procedures; compliance with regulations, guidelines, and accreditation requirements; employee health; direct intervention to prevent transmission of infectious diseases; education and training of healthcare workers; personnel resources; and nonpersonnel resources. The consensus panel used an evidence-based approach and categorized recommendations according to modifications of the scheme developed by the Clinical Affairs Committee of the Infectious Diseases Society of America and the Centers for Disease Control and Prevention's Hospital Infection Control Practices Advisory Committee.


Infectious cucumber mosaic virus RNA transcribed in vitro from clones obtained from cDNA amplified using the polymerase chain reaction.
Full-length cDNA to RNA 1, RNA 2 and RNA 3 of cucumber mosaic virus strain Q (CMV-Q) was amplified using the polymerase chain reaction (PCR). The first-strand primer contained a BamHI site and sequences complementary to the 3' terminus of the RNA. The second-strand primers contained a BamHI site, a T7 promoter and sequences corresponding to the 5' terminus of each RNA. After cleavage with BamHI, the PCR products were cloned into the BamHI site of the vector pEMBL9(+). Five clones of each RNA were selected and RNA transcripts were synthesized in vitro from each clone using T7 RNA polymerase. The constructs were designed to allow transcription to initiate precisely at the 5' terminus of each RNA. All the transcripts were found to be infectious when inoculated onto Nicotiana tabacum cv. Samsun plants in sets of three, corresponding to RNA 1, RNA 2 and RNA 3. Of the transcript sets, four induced symptoms indistinguishable from symptoms induced by CMV-Q RNAs. However a fifth transcript set induced much more severe symptoms. Plasmids were also constructed to allow synthesis of transcripts with one or two additional G residues at the 5' terminus of each RNA. Although the yields of such transcripts synthesized in vitro with T7 RNA polymerase were higher, their infectivity was lower than that of those with no additional residues at their 5' termini.


Detection and structural analysis of lipid-derived radicals in vitro and in vivo.
Lipids can be oxidized by reactive oxygen species, resulting in lipid peroxidation and the formation of reactive metabolites such as lipid-derived electrophiles. These products have been reported to induce inflammation, angiogenesis, and ferroptosis. Lipid peroxidation can produce many different products, each of which performs a different function, and which can be challenging to detect in vivo. The initial products of lipid oxidation are lipid-derived radicals, which can cause extensive chain reactions leading to lipid peroxidation. Hence, the ability to detect lipid radicals may provide information about this important class of molecules and the mechanism by which they cause cellular and tissue damage in a wide range of oxidative conditions. In this review, we report recent scientific advances in the detection of lipid-derived radicals in vitro and in cultured cells. We also introduce the possibility of visualization and structural analysis of lipid-derived radicals generated not only in in cells but also in animal tissue samples from oxidative disease models, using fluorescence-based lipid radicals' detection probes. We anticipate that the various innovative techniques summarized in this paper will be applied and further developed to clarify the role of lipid peroxidation in the pathogenesis of oxidative stress-associated diseases.


Good is better than excellent: bowel preparation quality and adenoma detection rates.
Inadequate bowel cleansing is associated with missed lesions, yet whether polyp and adenoma detection rates (PDR, ADR) increase at the highest levels of bowel cleanliness is unknown. To evaluate the association between bowel preparation quality by using the Boston Bowel Preparation Scale (BBPS) and PDR and ADR among colonoscopies with adequate preparation. Cross-sectional analysis. Boston Medical Center (BMC) and the Clinical Outcomes Research Initiative (CORI). Average-risk ambulatory patients attending screening colonoscopy with adequate bowel preparation defined as BBPS score ≥6. Colonoscopy. PDR and ADR stratified by BBPS score. Among the 3713 colonoscopies at BMC performed by 19 endoscopists, the PDR, ADR, and advanced ADR were 49.8%, 37.7%, and 6.0%, respectively. Among the 5532 colonoscopies in CORI performed by 85 endoscopists at 41 different sites, the PDR was 44.5%, and the PDR for polyps >9 mm (surrogate for advanced ADR) was 6.2%. The PDR associated with total BBPS scores of 6, 7, and 8 were higher than those associated with a BBPS score of 9 at BMC (BBPS 6, 51%; BBPS 7, 53%; BBPS 8, 52% vs BBPS 9, 46%; P = .002) and CORI (BBPS 6, 51%; BBPS 7, 48%; BBPS 8, 45% vs BBPS 9, 40%; P < .0001). This trend persisted after we adjusted for age, sex, and race and/or ethnicity and was observed for ADR and advanced ADR. PDR was higher among good compared with excellent preparations at BMC (odds ratio [OR] 1.3; 95% confidence interval [CI], 1.0-1.5) and CORI (OR 4.7; 95% CI, 3.1-7.1). Retrospective study. The PDR and ADR decreased at the highest levels of bowel cleanliness. Endoscopists finding a pristine bowel preparation should avoid a sense of overconfidence for polyp detection during the inspection phase of screening colonoscopy and still perform a careful evaluation for polyps. Furthermore, endoscopists expending additional effort to maximize cleansing of the bowel should never sacrifice on their inspection technique or inspection time.


[Severe neuropsychiatric disorder developing during busulfan-containing regimen for stem cell transplantation].
Here we describe two patients that required interruption of a busulfan (BU) containing conditioning regimen due to severe mental disorder before stem cell transplantation. The first patient was a 66-year-old man scheduled for unrelated peripheral blood stem cell transplantation with fludarabine/BU conditioning for myelodysplastic syndrome. He received 9.6 mg/kg BU and developed hallucinations that worsened the next day. BU was stopped on the final day, but the patient became comatose (grade 4). He recovered the next day. The second patient was a 69-year-old man scheduled for autologous peripheral blood stem cell transplantation with thiotepa (TT)/BU conditioning for cerebral nervous system relapse of mantle cell lymphoma. He received 12.8 mg/kg BU and developed hallucinations. His mental symptoms worsened on the next day, and thus administration was stopped on the second day of TT. His symptoms improved the next day. Both patients were over 65 years old, and their psychiatric symptoms worsened 1-2 days after the final dose of BU. Our findings suggest that BU may cause psychiatric disorders in elderly patients. When performing BU conditioning, it may be necessary to avoid azole antifungal medication and acetaminophen and to reduce the dose or perform therapeutic dose monitoring for elderly patients.


Comparative study: Endografine (diatrizoate), Vasurix polyvidone (acetrizoate), Dimer-X (iocarmate) and Hexabrix (ioxaglate) in hysterosalpingography.
Side effects of hysterosalpingography with Dimer-X, Hexabrix, Vasurix polyvidone and Endografine in 142 consecutive patients, receiving one of the four tested media were evaluated from replies to postal questionnaires. The Dimer-X group had a higher incidence of nausea and dizziness. The Endografine group had a higher incidence of abdominal pain. These differences occur especially in the age groups under 30 years. Hexabrix and Vasurix polyvidone are considered the best contrast media for hysterosalpingography and perhaps because of its low toxicity Hexabrix should be preferred.


Diethylstilbestrol and docetaxel: a Phase II study of tubulin active agents in patients with metastatic, androgen-independent prostate cancer.
The addition of diethylstilbestrol to docetaxel modified tubulin composition and improved the response of prostate cancer to chemotherapy in preclinical models. An attempt was made to recapitulate the observations in a clinical trial. Twenty-nine patients with progressive, metastatic, chemotherapy-naive androgen-independent prostate cancer were treated with diethylstilbestrol 1 mg daily and 5 mg on the day before docetaxel and docetaxel 36 mg/m(2) intravenously weekly for 3 weeks of a 4-week cycle. Prophylactic anticoagulation was used in all patients. Patients were assessed by prostate-specific antigen (PSA) monthly and computed tomography (CT) and bone scans every 3 cycles. The Response Evaluation Criteria in Solid Tumors (RECIST) criteria and PSA decline by >50% maintained for 4 weeks were used to assess activity. The median age was 68 years (range, 56-84 years), Southwest Oncology Group performance status 0 (score range, 0-2), alkaline phosphatase 120 U/L (range, 49-523), hemoglobin (Hgb) 12.6 g/dL (range, 9.2-16.3), PSA 66 ng/dL (range, 4-1962). The median number of cycles administered was 6. Soft tissue metastases were present in 51% of patients and bone metastases in 93%. Twenty-nine patients are evaluable for response. Of these, 20 patients (69%, 95% confidence interval [CI], 49%-85%) had a PSA decline of >50% and the PSA declined by >90% in 12 patients (41%, 95% CI, 23.1%-58.9%). Of 15 patients with measurable disease, 6 (40%, 95% CI, 23.5%-61%) had a partial response. Median time to progression was 6 months (range, 3-19 months). Fifteen patients (51%) suffered grade 3/4 toxicity. Two patients died of causes unrelated to therapy and another died from a steroid-induced ulcer. Six patients developed thrombosis and of those tested 75% had Factor V mutations. Pretreatment PSA, performance status, Hgb, and alkaline phosphatase had no impact on the likelihood of response. The combination of diethylstilbestrol and docetaxel produces a significant level of activity, measured by PSA decline and measurable disease response rate, and except for venous thrombosis the toxicity appears similar to that seen with docetaxel plus prednisone. These results suggest that tubulin modulation with diethylstilbestrol may improve the therapeutic efficacy of docetaxel and the combination is worthy of further study.


Fibronectin-mediated adhesion of fibroblasts: inhibition by dermatan sulfate proteoglycan and evidence for a cryptic glycosaminoglycan-binding domain.
Dermatan sulfate proteoglycans (DS-PGs) isolated from bovine articular cartilage have been examined for their effects on the adhesive responses of BALB/c 3T3 cells and bovine dermal fibroblasts on plasma fibronectin (pFN) and/or type I collagen matrices, and compared to the effects of the chondroitin sulfate/keratan sulfate proteoglycan monomers (CS/KS-PGs) from cartilage. DS-PGs inhibited the attachment and spreading of 3T3 cells on pFN-coated tissue culture substrata much more effectively than the cartilage CS/KS-PGs reported previously; in contrast, dermal fibroblasts were much less sensitive to either proteoglycan class unless they were pretreated with cycloheximide. Both cell types failed to adhere to substrata coated only with the proteoglycans; binding of the proteoglycans to various substrata has also been quantitated. While a strong inhibitory effect was obtained with the native intact DS-PGs, little inhibitory effect was obtained with isolated DS chains (liberated by alkaline-borohydride cleavage) or with core protein preparations (liberated by chondroitinase ABC digestion). In marked contrast, DS-PGs did not inhibit attachment or spreading responses of either 3T3 or dermal fibroblasts on type I collagen-coated substrata when the collagen was absorbed with pFN alone, DS-PGs alone, or the two in combination. These results support evidence for (a) collagen-dependent, fibronectin-independent mechanisms of adhesion of fibroblasts, and (b) different sites on the collagen fibrils where DS-PGs bind and where cell surface "receptors" for collagen bind. Experiments were developed to determine the mechanism(s) of inhibition. All evidence indicated that the mechanism using the intact pFN molecule involved the binding of the DS-PGs to the glycosaminoglycan (GAG)-binding sites of substratum-bound pFN, thereby inhibiting the interaction of the fibronectin with receptors on the cell surface. This was supported by affinity chromatography studies demonstrating that DS-PGs bind completely and effectively to pFN-Sepharose columns whereas only a subset of the cartilage CS/KS-PG binds weakly to these columns. In contrast, when a 120-kD chymotrypsin-generated cell-binding fragment of pFN (CBF which has no detectable GAG-binding activity as a soluble ligand) was tested in adhesion assays, DS-PGs inhibited 3T3 adherence on CBF more effectively than on intact pFN. A variety of experiments indicated that the mechanism of this inhibition also involved the binding of DS-PGs to only substratum-bound CBF due to the presence of a cryptic GAG-binding domain not observed in the soluble CBF.(ABSTRACT TRUNCATED AT 400 WORDS)


Chronic application of topiramate and carbamazepine differentially affects the EEG and penicillin-induced epileptiform activity in rats.
Carbamazepine (CBZ) and topiramate (TPM) are among commonly used antiepileptic drugs. The acute actions of these drugs are well known but the effects of long-term use on partially induced epileptiform characteristics are yet to be clarified. The rats were received CBZ (154 mg/kg/day), TPM (10 mg/kg/day) or tap water by gavage. We investigated penicillin-induced cortical epileptiform activity and electroencephalogram spectral power of rats by using electrocorticogram recordings. Animals were anesthetized with i.p. urethane (1·25 g/kg). Analysis of electroencephalogram recordings prior to epileptiform activity revealed that 3-week treatment of CBZ significantly increased relative power of delta (P<0·01) while reduced alpha (P<0·017) and beta (P<0·017) relative power compared to both control group and TPM group. TPM had no effect on absolute power and relative power of any frequency band. TPM treatment of 21 days significantly reduced the spike frequency (P<0·01). This preventive effect was missing in CBZ-treated rats. Upon the application of the last dose of drugs during ongoing epileptiform activity, the drugs suppressed the epileptiform activity. However, TPM was more rapid and effective than CBZ. In conclusion, our in vivo electrophysiological data suggest that TPM is more effective in animal model of partial epilepsy at the applied doses in this study.


Ghrelin and ghrelin receptor modulation of psychostimulant action.
Ghrelin (GHR) is an orexigenic gut peptide that modulates multiple homeostatic functions including gastric emptying, anxiety, stress, memory, feeding, and reinforcement. GHR is known to bind and activate growth-hormone secretagogue receptors (termed GHR-Rs). Of interest to our laboratory has been the assessment of the impact of GHR modulation of the locomotor activation and reward/reinforcement properties of psychostimulants such as cocaine and nicotine. Systemic GHR infusions augment cocaine stimulated locomotion and conditioned place preference (CPP) in rats, as does food restriction (FR) which elevates plasma ghrelin levels. Ghrelin enhancement of psychostimulant function may occur owing to a direct action on mesolimbic dopamine function or may reflect an indirect action of ghrelin on glucocorticoid pathways. Genomic or pharmacological ablation of GHR-Rs attenuates the acute locomotor-enhancing effects of nicotine, cocaine, amphetamine and alcohol and blunts the CPP induced by food, alcohol, amphetamine and cocaine in mice. The stimulant nicotine can induce CPP and like amphetamine and cocaine, repeated administration of nicotine induces locomotor sensitization in rats. Inactivation of ghrelin circuit function in rats by injection of a ghrelin receptor antagonist (e.g., JMV 2959) diminishes the development of nicotine-induced locomotor sensitization. These results suggest a key permissive role for GHR-R activity for the induction of locomotor sensitization to nicotine. Our finding that GHR-R null rats exhibit diminished patterns of responding for intracranial self-stimulation complements an emerging literature implicating central GHR circuits in drug reward/reinforcement. Finally, antagonism of GHR-Rs may represent a smoking cessation modality that not only blocks nicotine-induced reward but that also may limit weight gain after smoking cessation.


Campylobacter and Salmonella are prevalent in broiler farms in Kyushu, Japan: results of a 2-year distribution and circulation dynamics audit.
To elucidate the distribution and circulation dynamics of Campylobacter and Salmonella in Japanese chicken broiler flocks. A 2-year investigation of the distribution of Campylobacter and Salmonella was conducted in 25 broiler flocks at nine farms in Japan from 2013 to 2014. Campylobacter and Salmonella tested positive in 11 (44·0%) and 24 (96·0%) broiler flocks respectively. One hundred and ninety-five Campylobacter and 184 Salmonella isolates were characterized into 12 Campylobacter (including two novel genotypes) and three Salmonella MLST genotypes. Only Salmonella isolation between caecal and environmental samples were significantly correlated. Further, one litter sample tested positive for Salmonella before new chicks were introduced. The Campylobacter strains rapidly lost culturability within 2-18 days; in contrast, the Salmonella strains survived from 64-211 days in artificially inoculated water samples. No persistent circulation-mediated Campylobacter contamination was observed. In contrast, circulation of Salmonella in broiler houses was seen, apparently due to the litter excreted from broiler flocks, as well as Salmonella-contaminated water and feed. This paper provides the distribution, genotypic data and circulation dynamics of Campylobacter and Salmonella as recently observed in Japanese chicken broiler farms.


Treatment of Periimplantitis with Electrolytic Cleaning versus Mechanical and Electrolytic Cleaning: 18-Month Results from a Randomized Controlled Clinical Trial.
Aim of the study: This RCT assesses patients' 18-month clinical outcomes after the regenerative therapy of periimplantitis lesions using either an electrolytic method (EC) to remove biofilms or a combination of powder spray and an electrolytic method (PEC). Materials and Methods: Twenty-four patients (24 implants) suffering from periimplantitis were randomly treated by EC or PEC followed by augmentation and submerged healing. Probing pocket depth (PPD), Bleeding on Probing (BoP), suppuration, and standardized radiographs were assessed before surgery (T0), 6 months after augmentation (T1), and 6 (T2) and 12 (T3) months after the replacement of the restoration. Results: The mean PPD changed from 5.8 ± 1.6 mm (T0) to 3.1 ± 1.4 mm (T3). While BoP and suppuration at T0 were 100%, BoP decreased at T2 to 36.8% and at T3 to 35.3%. Suppuration was found to be at a level of 10.6% at T2 and 11.8% at T3. The radiologic bone level measured from the implant shoulder to the first visible bone to the implant contact was 4.9 ± 1.9 mm at mesial sites and 4.4 ± 2.2 mm at distal sites at T0 and 1.7 ± 1.7 mm and 1.5 ± 17 mm at T3. Conclusions: Significant radiographic bone fill and the improvement of clinical parameters were demonstrated 18 months after therapy.


Remote monitoring of implantable cardiac monitors in patients with unexplained syncope: Predictors of false-positive alert episodes.
Remote monitoring is recommended for patients with implantable cardiac monitors (ICMs), but compared to other cardiac implantable devices, ICMs are less accurate and transmit a higher number of alerts. The aim of this study was to investigate the predictors of false-positive (FP) arrhythmic alerts in patients with unexplained syncope who were implanted with ICM and followed by an automatic remote monitoring system. We retrospectively evaluated all consecutive patients who received a long-sensing vector ICM for unexplained syncope between January 2019 to September 2021 at our Syncope Unit. The primary endpoint was the incidence of the first FP episode. The secondary endpoints included assessing the incidence of FP episodes for all types of algorhythms and indentifying the reasons for the misdetection of these episodes. Among 105 patients (44.8% males, median age 51 years), 51 (48.6%) transmitted at least one FP alert during a median follow-up of 301 days. The presence of pre-ventricular complexes (PVCs) on the resting electrocardiogram was the only clinical characteristic associated with an increased risk of FP alerts (adjusted Hazard ratio [HR] 5.76 [2.66-12.4], p = 0.010). The other significant device-related variables were a low-frequency filter at 0.05 Hz versus the default 0.5 Hz (adjusted HR 3.82 [1.38-10.5], p = 0.010) and the R-wave amplitude (adjusted HR 0.35 [0.13-0.99], p = 0.049). Patients who have PVCs are at higher risk of inappropriate ICM activations. To reduce the occurrence of FP alerts, it may be beneficial to target a large R-wave amplitude during device insertion and avoid programming a low-frequency filter at 0.05 Hz.


Thin-disk ring amplifier for high pulse energy.
A CPA laser system for high pulse energy at high average power has been developed. The system is based on Yb:YAG thin-disk technology. It provides two laser beams with more than 500 mJ pulse energy each at 100 Hz repetition rate and 2 ps pulse duration. The system consists of a common oscillator, a grating stretcher and compressor and two identical amplifier chains that are both equipped with a regenerative amplifier and a ring amplifier. The compressor supports an individual alignment of the dispersion for the two laser channels.


An investigation of trauma-centered inpatient treatment for adult survivors of abuse.
The purpose of this study was to examine a comprehensive inpatient treatment program designed for adult survivors of childhood abuse with posttraumatic stress disorder (PTSD). One hundred and thirty-two formerly abused individuals completed clinician-administered and self-administered measures of PTSD symptomatology at admission and discharge. All participants experienced a range of physical, sexual, and/or emotional abuse as children prior to the age of 17. Approximately one-third of these individuals also completed measures at 3-months postdischarge and 1-year postdischarge. Data were collected using a clinician-administered PTSD measure and self-administered PTSD measure at admission and discharge. On admission, all participants met criteria for a diagnosis of PTSD. Analyses revealed that the program was effective in reducing symptoms from admission to discharge. Additionally, treatment gains were maintained at 1-year postdischarge. The findings of this investigation suggest that the current intensive inpatient group treatment program appears to reduce PTSD symptoms effectively for a sample of adult survivors of abuse.


Purification, concentration, and inactivation of Venezuelan equine encephalitis virus.
Venezuelan equine encephalitis (VEE) virus was purified and concentrated by chromatography of tissue culture supernatant fluids on diethylaminoethyl-cellulose columns. Stepwise gradient elution studies indicated a broad elution pattern for the virus, with recovery occurring from 0.05 to 0.7 m NaCl. Optical density, infectivity, hemagglutination (HA), and complement fixation (CF) assays indicated that complete recovery of input virus in highly purified form was possible. Single-step elution with 0.7 m tris(hydroxymethyl)aminomethane-succinate-salt buffer resulted in a virus volume decrease of 85% with a concomitant increase in infectivity and antigenicity. Recoveries consistently equaled or exceeded 100% of the input preparations. Additional purification of column-recovered virus was obtained by sedimentation of pooled virus eluates on 50% sucrose cushions. Exposure of borate saline and 0.5% histidine suspensions of purified VEE virus preparations to 6 x 10(6) r of gamma radiation resulted in a loss of infectivity for tissue culture and a loss of lethality for weanling and suckling mice. Inactivation was an exponential function of the dosage. In contrast to infectivity, antigencity (HA and CF) of both saline and histidine preparations was retained after irradiation with doses as high as 6 x 10(6) r. Purified and irradiated VEE virus preparations have been successfully used for routine serological tests and are being evaluated as vaccines.


A public health hazard mitigation planning process.
The Texas A&M Health Science Center School of Rural Public Health, a member of the Training and Education Collaborative System Preparedness and Emergency Response Learning Center (TECS-PERLC), has long-standing partnerships with 2 Health Service Regions (Regions) in Texas. TECS-PERLC was contracted by these Regions to address 2 challenges identified in meeting requirements outlined by the Risk-Based Funding Project. First, within Metropolitan Statistical Areas, there is not a formal authoritative structure. Second, preexisting tools and processes did not adequately satisfy requirements to assess public health, medical, and mental health needs and link mitigation strategies to the Public Health Preparedness Capabilities, which provide guidance to prepare for, respond to, and recover from public health incidents. TECS-PERLC, with its partners, developed a framework to interpret and apply results from the Texas Public Health Risk Assessment Tool (TxPHRAT). The 3-phase community engagement-based TxPHRAT Mitigation Planning Process (Mitigation Planning Process) and associated tools facilitated the development of mitigation plans. Tools included (1) profiles interpreting TxPHRAT results and identifying, ranking, and prioritizing hazards and capability gaps; (2) a catalog of intervention strategies and activities linked to hazards and capabilities; and (3) a template to plan, evaluate, and report mitigation planning efforts. The Mitigation Planning Process provided a framework for Regions to successfully address all funding requirements. TECS-PERLC developed more than 60 profiles, cataloged and linked 195 intervention strategies, and developed a template resulting in 20 submitted mitigation plans. A public health-focused, community engagement-based mitigation planning process was developed by TECS-PERLC and successfully implemented by the Regions. The outcomes met all requirements and reinforce the effectiveness of academic practice partnerships and importance of community engagement in mitigation planning. Additional funding has been approved to expand the Mitigation Planning Process to all counties in Texas with local health departments.


Non-parametric algorithmic generation of neuronal morphologies.
Generation algorithms allow for the generation of Virtual Neurons (VNs) from a small set of morphological properties. The set describes the morphological properties of real neurons in terms of statistical descriptors such as the number of branches and segment lengths (among others). The majority of reconstruction algorithms use the observed properties to estimate the parameters of a priori fixed probability distributions in order to construct statistical descriptors that fit well with the observed data. In this article, we present a non-parametric generation algorithm based on kernel density estimators (KDEs). The new algorithm is called KDE-NEURON: and has three advantages over parametric reconstruction algorithms: (1) no a priori specifications about the distributions underlying the real data, (2) peculiarities in the biological data will be reflected in the VNs, and (3) ability to reconstruct different cell types. We experimentally generated motor neurons and granule cells, and statistically validated the obtained results. Moreover, we assessed the quality of the prototype data set and observed that our generated neurons are as good as the prototype data in terms of the used statistical descriptors. The opportunities and limitations of data-driven algorithmic reconstruction of neurons are discussed.


Oxidation and base-catalyzed elimination of the saccharide portion of GSLs having very different polarities.
Glycosphingolipids (GSLs), present in cell membranes, participate in a variety of biological functions. Although their exact role(s) may not be understood, it has been shown that 1) embryos lacking glucosylceramide synthase activity do not develop normally, 2) GSLs can affect neuritogenesis, and 3) they can function as receptors for some pathogens. To study the role of the saccharide portion of a GSL in any of these functions, it is necessary to either isolate it from the intact GSL or synthesize it. Because syntheses are more complex, modifications were made to the oxidation/elimination procedure previously described for the isolation of the saccharide portion of GM1 and GD1a to enable it to be used with GSLs of varying polarity. The key is to use a mixture of GSLs that differ in polarity. This appears to eliminate problems encountered when purified GSLs such as sulfatide or GT1b are used.


Chronobiological studies on body search, oviposition and emergence of Megaselia scalaris (Diptera, Phoridae) in controlled conditions.
Circadian clocks have evolved to synchronize physiology, metabolism and behaviour to the 24-h geophysical cycles of the Earth. Understanding the circadian clock mechanism could play an important role in forensic entomology because it temporally gates behaviour such as locomotor activities, feeding, mating, egg laying and adult emergence which could provide useful information for crime reconstruction. The scuttle fly Megaselia scalaris colonises both exposed and buried bodies, whether indoors or outdoors. Locomotor activity, oviposition and adult emergence of this species have been investigated using technologies employed in many previous Drosophila circadian studies. The results reported here clearly highlight the underlying role of the circadian clock in regulating the behaviour of males and females of M. scalaris, and show the role of light as a "zeitgeber" for clock resetting. In contrast to Calliphoridae, M. scalaris can reach the oviposition site and lay eggs in darkness both during the day and the night, although the number of ovipositing females is lower under subjective darkness. The number of eggs laid shows a clear circadian rhythm with much higher numbers laid during the day than during night or subjective night. In conclusion, locomotor activity and oviposition rate of M. scalaris is under circadian clock control with significant forensic implications.


The effect of an oxygenating agent upon recurrent aphthous stomatitis -- a double-blind clinical trial.
Fifty subjects enrolled in a double-blind clinical trial to test the effectiveness of a solution of carbamide peroxide in 10% glycerol upon healing time and pain relief of recurrent aphthous stomatitis. No apparent difference was experienced in either duration of healing or pain relief between the 25 subjects who received the active medication and the 25 subjects who received a placebo.


[Characteristics of Actinomyces griseus actinophages and the production of phage-resistant mutants of the producer of grisin].
Analysis of phagolysates in production of fodder grisin provided determination of 3 serologically and morphologically different actinophages, 2 of which, i. e. Pg 81 and Pg 100 affected the sensitive culture as virulent phages and 1, i. e. Pg 2 affected it as a moderate phage. Investigation of the fermentation broth of strain 15 of Act. griseus did not confirm the supposition that this strain was polylysogenic and actinophages Pg 81 and Pg 100 were virulent mutants of moderate phages present in the cells. A phenomenon of limitation and modification controlled by the host cell was noted in the sensitive culture infected with actinophage Pg 81. The actinomycete mutants which did not limit but modified the phage were isolated. Under the effect of chemical mutagens the actinomycete mutants stable to virulent phages were selected. Treatment of the phage-stable mutants with ethylene imine and UV-light provided an increase in their antibiotic production levels by 75 to 80 per cent.


Iron photoreduction and oxidation in an acidic mountain stream.
In a small mountain stream in Colorado that receives acidic mine drainage, photoreduction of ferric iron results in a well-defined increase in dissolved ferrous iron during the day. To quantify this process, an instream injection of a conservative tracer was used to measure discharge at the time that each sample was collected. Daytime production of ferrous iron by photoreduction was almost four times as great as nighttime oxidation of ferrous iron. The photoreduction process probably involves dissolved or colloidal ferric iron species and limited interaction with organic species because concentrations of organic carbon are low in this stream.


A Longitudinal Approach to Changes in the Motivation of Dutch Pharmacists in the Current Continuing Education System.
Objective. To explore the changes in motivation of Dutch pharmacists for Continuing Education (CE) in the Dutch CE system. Methods. Pharmacists' motivation was measured across three time points with the Academic Motivation Scale, based on the Self-Determination Theory of motivation. The Latent Growth Modelling technique was used to analyze these data. Results. Over a period of 21 months, Controlled Motivation had increased and Relative Autonomous Motivation of Dutch pharmacists had decreased. Traineeship was the only demographic factor with a significant influence on the change in motivation. No subgroups with different trajectories could be identified. Conclusion. Relative Autonomous Motivation of Dutch pharmacists for CE decreases over time. This indicates a loss of Autonomous Motivation ("good" motivation) in favor of Controlled Motivation ("bad" motivation). Further research needs to be conducted to gain a better understanding of the association between pharmacist motivation and the features of the current CE system.


Rapid Eye Movement Sleep Behavior Disorder and Neurodegenerative Diseases: An Update.
Rapid eye movement sleep behavior disorder (RBD) is a sleep behavior disorder characterized by abnormal behaviors and loss of muscle atonia during rapid eye movement (REM) sleep. RBD is generally considered to be associated with synucleinopathies, such as Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), and usually precedes years before the first symptom of these diseases. It is believed that RBD predicts the neurodegeneration in synucleinopathy. However, increasing evidences have shown that RBD is also found in non-synucleinopathy neurodegenerative diseases, including Alzheimer's disease (AD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), etc. Sleep disturbance such as RBD may be an early sign of neurodegeneration in these diseases, and also serve as an assessment of cognitive impairments. In this review, we updated the clinical characteristics, diagnosis, and possible mechanisms of RBD in neurogenerative diseases. A better understanding of RBD in these neurogenerative diseases will provide biomarkers and novel therapeutics for the early diagnosis and treatment of the diseases.


Colobops: a juvenile rhynchocephalian reptile (Lepidosauromorpha), not a diminutive archosauromorph with an unusually strong bite.
Correctly identifying taxa at the root of major clades or the oldest clade-representatives is critical for meaningful interpretations of evolution. A small, partially crushed skull from the Late Triassic (Norian) of Connecticut, USA, originally described as an indeterminate rhynchocephalian saurian, was recently named Colobops noviportensis and reinterpreted as sister to all remaining Rhynchosauria, one of the earliest and globally distributed groups of herbivorous reptiles. It was also interpreted as having an exceptionally reinforced snout and powerful bite based on an especially large supratemporal fenestra. Here, after a re-analysis of the original scan data, we show that the skull was strongly dorsoventrally compressed post-mortem, with most bones out of life position. The cranial anatomy is consistent with that of other rhynchocephalian lepidosauromorphs, not rhynchosaurs. The 'reinforced snout' region and the 'exceptionally enlarged temporal region' are preservational artefacts and not exceptional among clevosaurid rhynchocephalians. Colobops is thus not a key taxon for understanding diapsid feeding apparatus evolution.


Improvement in facial aesthetics of orthognathic patients after surgery-first approach.
Facial appearance significantly affects psychosocial wellbeing, and an improvement in facial aesthetics is considered an essential outcome of orthognathic treatment. The surgery-first approach (SFA) has emerged as a promising alternative to the conventional orthodontics-first approach (OFA) due to its potential advantages in reducing treatment duration and cost, delivering early aesthetic improvement, and increasing patient satisfaction. However, its impact on final facial aesthetics and how it compares with the OFA has, to our knowledge, not yet been investigated. This retrospective study aimed to compare the improvement in facial aesthetics after orthognathic surgery in an SFA and an OFA group. Preoperative and postoperative 3-dimensional stereophotogrammetry facial images of 40 patients were evaluated by five professional assessors using the Global Aesthetic Improvement Scale (GAIS). Similar aesthetic improvement outcomes were found in both the SFA and OFA groups. The GAIS score significantly correlated with the following facial variables: upper lip projection, chin prominence, facial proportions, paranasal hollowing, lip competence, mandibular projection, and facial profile. No significant correlation was found between a change in aesthetic score and the surgical variables. There was a positive association between overall GAIS score and the gender and experience level of the individual assessors. This study suggests that aesthetic facial improvement achieved with the SFA is satisfactory and comparable to that of the OFA.


The search for efficient diagnostic and prognostic biomarkers of heart failure.
Several biomarkers have been tested for screening, diagnosis and prognosis purposes, as well as to guide treatment in heart failure, but only the assay of circulating B-type natriuretic peptides has widely recognized applications for clinical decision-making. Natriuretic peptides are sensitive in detecting the clinically overt or subclinical myocardial damage, but their plasma levels are increased following every generic insult to the cardiovascular system. Novel biomarkers are required to identify specific pathways of disease progression, such as diverse neurohormonal axes activation, inflammation and fibrogenesis, and to act as a tool for therapeutic tailoring. In this view, Gal-3 and ST-2 assays seem very promising, given their involvement in mechanisms of cardiac fibrosis and their prognostic value.


Interactions between pH, reactive species, and cells in plasma-activated water can remove algae.
Lightning strikes cause nitrogen to dissolve in water and form reactive nitrogen and oxygen species, which form natural fertilizers that can be absorbed through plant roots. Such processes during rainstorm events can be simulated by applying plasma to a solution. Plasma-activated water (PAW) has great potential as a source of various dissolved reactive chemical species. Different mixtures of species are produced using different solution compositions. Here, basil seeds were grown in PAW to prevent blooms of Chlorella vulgaris and ion chromatography and UV-vis spectroscopy were used to quantify reactive ions. NO2 -, NO3 -, and H2O2 were found to be key to the antialgal effect. Secondary reactive ions such as peroxynitrite (ONOO-, ONOOH) were also involved. The antialgal effect was strongly related to the pH around the algal cells. Acidification was predominantly caused by the generation of NO2 - and H2O2. After two weeks monitoring basil growth, the antifungal properties were preserved, few reactive oxygen species formed in the plasma zone, and only reactive nitrogen species were transformed into reactive peroxynitrite ions. The pH around the cells was determined using an iridium oxide microelectrode. The PAW antialgal mechanism depended on acidic conditions (pH 2.2, at which peroxynitrite can be generated) under which ONOOH penetrated the algal cell membranes, destroying the cells and preventing growth. This practical and sustainable PAW process allows a surprising amount of fertilizer to be generated with an antialgal effect that could be used in various eco-friendly agricultural processes under ambient conditions.


Lipoprotein lipase reaches the capillary lumen in chickens despite an apparent absence of GPIHBP1.
In mammals, GPIHBP1 is absolutely essential for transporting lipoprotein lipase (LPL) to the lumen of capillaries, where it hydrolyzes the triglycerides in triglyceride-rich lipoproteins. In all lower vertebrate species (e.g., birds, amphibians, reptiles, fish), a gene for LPL can be found easily, but a gene for GPIHBP1 has never been found. The obvious question is whether the LPL in lower vertebrates is able to reach the capillary lumen. Using purified antibodies against chicken LPL, we showed that LPL is present on capillary endothelial cells of chicken heart and adipose tissue, colocalizing with von Willebrand factor. When the antibodies against chicken LPL were injected intravenously into chickens, they bound to LPL on the luminal surface of capillaries in heart and adipose tissue. LPL was released rapidly from chicken hearts with an infusion of heparin, consistent with LPL being located inside blood vessels. Remarkably, chicken LPL bound in a specific fashion to mammalian GPIHBP1. However, we could not identify a gene for GPIHBP1 in the chicken genome, nor could we identify a transcript for GPIHBP1 in a large chicken RNA-seq data set. We conclude that LPL reaches the capillary lumen in chickens - as it does in mammals - despite an apparent absence of GPIHBP1.


Fluorescence-based fibre optic arrays: a universal platform for sensing.
Optical fibres provide a universal sensing platform as they are easily integrated with a multitude of different sensing schemes. Such schemes enable the preparation of a multitude of sensors from relatively straightforward pH sensors, to more complex ones, including artificial olfaction sensors, high-density oligonucleotide arrays, and high-throughput cell-based arrays. Imaging fibre bundles comprised of thousands of fused optical fibres are the basis for an optically connected, individually addressable parallel sensing platform. Fibre optic imaging bundles possess miniature feature sizes (3-10 micron diameter fibres), allowing high-density sensor packing (approximately 2 x 10(7) sensors per cm2). Imaging fibre bundles transmit coherent images enabling combined imaging and sensing, relating the responses monitored by the sensor to observable physical changes. The individual fibre cores can also be selectively etched to form a high-density microwell array capable of housing complementary sized microsensors. The miniature feature sizes facilitate a faster response and more sensitive measurement capabilities. The platform is extremely versatile in its sensing design, allowing the sensing scheme to be tailored to fit the experimental design, whether for monitoring single analytes or more complex multiplexed assays. A number of sensing schemes and applications are described in this review.


Cost-management solutions for health care.
Employers do not need the new math to compute that health care reform equals increased costs. Faced with this dilemma, they are exploring cost-management solutions to keep up with reform and at the same time keep down costs.


HypertenGene: extracting key hypertension genes from biomedical literature with position and automatically-generated template features.
The genetic factors leading to hypertension have been extensively studied, and large numbers of research papers have been published on the subject. One of hypertension researchers' primary research tasks is to locate key hypertension-related genes in abstracts. However, gathering such information with existing tools is not easy: (1) Searching for articles often returns far too many hits to browse through. (2) The search results do not highlight the hypertension-related genes discovered in the abstract. (3) Even though some text mining services mark up gene names in the abstract, the key genes investigated in a paper are still not distinguished from other genes. To facilitate the information gathering process for hypertension researchers, one solution would be to extract the key hypertension-related genes in each abstract. Three major tasks are involved in the construction of this system: (1) gene and hypertension named entity recognition, (2) section categorization, and (3) gene-hypertension relation extraction. We first compare the retrieval performance achieved by individually adding template features and position features to the baseline system. Then, the combination of both is examined. We found that using position features can almost double the original AUC score (0.8140 vs.0.4936) of the baseline system. However, adding template features only results in marginal improvement (0.0197). Including both improves AUC to 0.8184, indicating that these two sets of features are complementary, and do not have overlapping effects. We then examine the performance in a different domain--diabetes, and the result shows a satisfactory AUC of 0.83. Our approach successfully exploits template features to recognize true hypertension-related gene mentions and position features to distinguish key genes from other related genes. Templates are automatically generated and checked by biologists to minimize labor costs. Our approach integrates the advantages of machine learning models and pattern matching. To the best of our knowledge, this the first systematic study of extracting hypertension-related genes and the first attempt to create a hypertension-gene relation corpus based on the GAD database. Furthermore, our paper proposes and tests novel features for extracting key hypertension genes, such as relative position, section, and template features, which could also be applied to key-gene extraction for other diseases.


Ultrasound-guided perineural steroid injection to treat intractable pain due to sciatic nerve injury.
Sciatic neuropathy is a rare but serious complication of cardiac surgery. Neuropathic pain following nerve injury can be severely debilitating and largely resistant to treatment. We present a case of this complication where ultrasound-guided perineural steroid injection at the site of the sciatic nerve injury provided excellent pain relief and facilitated subsequent rehabilitation. A 17-yr-old boy developed bilateral sciatic neuropathy after a nine-hour cardiac surgical procedure in the supine position, resulting in debilitating dysesthesia refractory to neuropathic pain therapies and leading to severe functional limitation. With magnetic resonance imaging of the lower extremities, the location of the lesion was determined to be from the level of the superior gemellus to the level of the quadratus femoris. An ultrasound-guided injection of triamcinolone 20 mg and lidocaine 40 mg around both sciatic nerves at the level of the lesion was administered two months after the surgery, and the pain score (rated on a scale 0-10) at rest decreased from 9-10 to 1 two weeks after the injection. There are a limited number of reports in the literature on sciatic nerve injuries associated with cardiac surgery. This case illustrates the efficacy of ultrasound-guided steroid injection around sciatic nerves at the level of superior gemellus in treating our patient's neuropathic pain.


The psychosocial meanings of living related kidney organ donation: recipient and donor perspectives--literature review.
There is a shortage of information about the psychosocial influences that impact family decision making for a living related donor kidney transplant. This article reviews research findings related to the psychosocial meanings of donation. potential recipient's decisions about the kidney donor, and families' roles and expectations. The social meaning of organ donation as a "gift" and family dynamics involved in decision making are addressed. The findings show that families make decisions about kidney donation in different ways. Emotional responses of potential kidney recipients include ambivalence, passivity, and denial of the severity of their illness. Emotional responses of potential family donors range from unrestrained willingness to ambivalence and concerns of coercion. Improved accuracy of health assessment and awareness of the importance of psychosocial perspectives is needed. Assessment questions addressing the fears and concerns of potential recipient and donor are suggested.


Quantifying the literature of computer-aided instruction in medical education.
To characterize the literature about computer-aided instruction (CAI) as it relates to medical education. A descriptive study using the Medline and ERIC databases, reviewing articles pertaining to CAI for medical professionals. The literature was evaluated with respect to type of article, journal, language, and year of publication. The search yielded 2,840 citations, 92% of which were in English. The number of journals with at least one citation was 747. Less than 5% of the 5,147 authors had three or more articles published in the CAI literature. Of the citations with abstracts, 60% were demonstrations of a CAI application, 11% were media-comparative studies, and 13% were analyses of the CAI field. While the pace of article publication increased markedly over time, the percentages of article types did not change significantly over time. Less than 10% of CAI articles appeared in core medical journals. Medical CAI is an increasingly popular topic of research and publication. However, these studies appear in journals with smaller circulations, are predominantly demonstration articles, and are generally written by authors with two or fewer publications. Evaluation articles remain less common. A series of analytic articles has appeared offering substantive suggestions for better research design. These suggestions appear to have gone unheeded. CAI investigators need be more aware of the gaps in the existing body of CAI publications, and the inherent difficulties of this type of research, if this literature is to move beyond this early stage of development.


Glandular lesions of the cervix. Validity of cytologic criteria used to differentiate reactive changes, glandular intraepithelial lesions and adenocarcinoma.
To determine the validity and reproducibility of various cytologic criteria as discriminators between different glandular lesions of the cervix. Seventy-three cervicovaginal smears with glandular lesions and a documented histologic and/ or clinical correlation were studied. The lesions included reactive changes, low grade glandular intraepithelial lesion (LGIL) (encompassing endocervical glandular dysplasia), high grade glandular intraepithelial lesion (HGIL) (encompassing adenocarcinoma in situ) and invasive adenocarcinoma (IA). Twenty-three cytologic criteria were used to evaluate the smears, and the results were scored and statistically analyzed. Reactive lesions consistently showed well-defined cell borders, normal nuclear/cytoplasmic (N/C) ratio, minimal or absent nuclear overlapping, round to oval nuclei with fine chromatin and prominent nucleoli. HGIL (adenocarcinoma in situ) showed feathered edges, rosettes, cell strips, increased N/C ratio, elongated nuclei, marked nuclear overlapping and nuclei with hyperchromatic, coarse chromatin. IA shared features with HGIL but had a greater tendency to show a dirty background, single cells, mitotic figures, nuclear pleomorphism and large nucleoli. Logistic regression studies indicated that the presence of mitotic figures, a dirty background and single cells increases the odds ratio for predicting invasion. LGIL showed features similar to those of high grade lesions, but the changes were more subtle. These lesions were also less cellular and less likely to show cell strips, feathered edges and rosettes. Similar to HGIL and IA, LGIL showed nuclear overlapping, increased N/C ratio, oval to elongated nuclei and nuclear hyperchromasia. There is overlap between the cytologic criteria for the various glandular lesions of the cervix. However, some of these criteria can consistently distinguish clinically significant lesions from reactive benign changes seen in the glandular epithelium.


Multiplex H2 coherent anti-Stokes Raman scattering thermometry with a modeless laser.
The use of a modeless laser as the Stokes source for multiplex coherent anti-Stokes Raman scattering in molecular hydrogen is reported. The elimination of noise associated with mode competition in conventional standing wave lasers is shown to result in reliable and accurate single-shot thermometry of H(2) in a microwave-assisted diamond chemical vapor deposition plasma reactor. Single-shot temperatures are recorded with a precision of 7.3%. Possible improvements to this precision are discussed and applications of the technique for on-line process monitoring are briefly presented.


Transcriptomic analyses of xylan degradation by Prevotella bryantii and insights into energy acquisition by xylanolytic bacteroidetes.
Enzymatic depolymerization of lignocellulose by microbes in the bovine rumen and the human colon is critical to gut health and function within the host. Prevotella bryantii B(1)4 is a rumen bacterium that efficiently degrades soluble xylan. To identify the genes harnessed by this bacterium to degrade xylan, the transcriptomes of P. bryantii cultured on either wheat arabinoxylan or a mixture of its monosaccharide components were compared by DNA microarray and RNA sequencing approaches. The most highly induced genes formed a cluster that contained putative outer membrane proteins analogous to the starch utilization system identified in the prominent human gut symbiont Bacteroides thetaiotaomicron. The arrangement of genes in the cluster was highly conserved in other xylanolytic Bacteroidetes, suggesting that the mechanism employed by xylan utilizers in this phylum is conserved. A number of genes encoding proteins with unassigned function were also induced on wheat arabinoxylan. Among these proteins, a hypothetical protein with low similarity to glycoside hydrolases was shown to possess endoxylanase activity and subsequently assigned to glycoside hydrolase family 5. The enzyme was designated PbXyn5A. Two of the most similar proteins to PbXyn5A were hypothetical proteins from human colonic Bacteroides spp., and when expressed each protein exhibited endoxylanase activity. By using site-directed mutagenesis, we identified two amino acid residues that likely serve as the catalytic acid/base and nucleophile as in other GH5 proteins. This study therefore provides insights into capture of energy by xylanolytic Bacteroidetes and the application of their enzymes as a resource in the biofuel industry.


Features predicting the success of computerized decision support for prescribing: a systematic review of randomized controlled trials.
Computerized decision support systems (CDSS) are believed to have the potential to improve the quality of health care delivery, although results from high quality studies have been mixed. We conducted a systematic review to evaluate whether certain features of prescribing decision support systems (RxCDSS) predict successful implementation, change in provider behaviour, and change in patient outcomes. A literature search of Medline, EMBASE, CINAHL and INSPEC databases (earliest entry to June 2008) was conducted to identify randomized controlled trials involving RxCDSS. Each citation was independently assessed by two reviewers for outcomes and 28 predefined system features. Statistical analysis of associations between system features and success of outcomes was planned. Of 4534 citations returned by the search, 41 met the inclusion criteria. Of these, 37 reported successful system implementations, 25 reported success at changing health care provider behaviour, and 5 noted improvements in patient outcomes. A mean of 17 features per study were mentioned. The statistical analysis could not be completed due primarily to the small number of studies and lack of diversity of outcomes. Descriptive analysis did not confirm any feature to be more prevalent in successful trials relative to unsuccessful ones for implementation, provider behaviour or patient outcomes. While RxCDSSs have the potential to change health care provider behaviour, very few high quality studies show improvement in patient outcomes. Furthermore, the features of the RxCDSS associated with success (or failure) are poorly described, thus making it difficult for system design and implementation to improve.


Treatment persistence and maintenance dose titration among ulcerative colitis patients on biologics: a pooled study of three United States claim databases.
This real-world study evaluated biologic treatment patterns in patients with moderate-to-severe ulcerative colitis (UC). IQVIA PharMetrics, IBM MarketScan, and Optum Clinformatics were pooled to identify UC patients with ≥1 claim for UC and ≥1 claim for adalimumab (ADA), golimumab (GOL), infliximab (IFX), or vedolizumab (VDZ). The index date for each biologic was the first claim for that biologic. Patients could be included in >1 cohort if they switched biologics during the identification period. Continuous eligibility for medical/pharmacy benefits was required for 12 months before (baseline) and after (follow-up) the index date. Patients lacking claims for any biologic during baseline were categorized as bio-naïve; those with any biologic claim were categorized as bio-experienced. Persistence was defined as the proportion of patients that remained on the index biologic without a gap between claims of >28 days for ADA, >56 days for GOL, and >112 days for IFX and VDZ. Dose titration was assessed among patients with ≥2 maintenance doses during follow-up among ADA, GOL, and VDZ patients. In total, 6,106 bio-naïve UC patients and 1,027 bio-experienced UC patients were identified. Patients treated with VDZ and IFX had the highest persistence followed by ADA and GOL patients for bio-naïve and bio-experienced, respectively. ADA patients had a numerically higher proportion of patients with 50% dose escalation, followed by VDZ and GOL; 50% dose reduction was observed in ≤1% patients. In this descriptive study of UC patients without confounder adjustment, VDZ persistence was numerically highest followed by IFX, GOL, and ADA across both populations.


Investigation of CO2 Photoreduction in an Annular Fluidized Bed Photoreactor by MP-PIC Simulation.
Carbon dioxide (CO2) photoreduction is a promising process for both mitigating CO2 emissions and providing chemicals and fuels. A gas-solid two-phase annular fluidized bed photoreactor (FBPR) would be preferred for this process due to its high mass-transfer rate and easy operation. However, CO2 photoreduction using the FBPR has not been widely researched to date. The Lagrangian multiphase particle-in-cell (MP-PIC) simulation with computational fluid dynamic models is a new and robust approach to explore the multiphase reaction system in the gas-solid fluidized bed. Therefore, the purpose of this paper is to investigate CO2 photoreduction in the FBPR by MP-PIC modeling to understand the intrinsic mechanism of solid flow, species mass transfer, and CO2 photoreaction. The MP-PIC models for solid flow in the FBPR were validated by the bed expansion height and bubble size. The results showed the particle stress of the Lun model, the drag of the Ergun-WenYu (Gidaspow) model, and the coefficient of restitution e = 0.95 with the wall parameters e w = 0.9 and μw = 0.6 are the best fit to the experimental empirical correlations. The MP-PIC models developed in this work proved to be better than the Eulerian two-fluid modeling in the prediction of the bed expansion height and bubble size. It was also found from the simulation results that the maximum radiation intensity is in the half reactor height area, and the photocatalytic reaction mainly occurred around the inner wall. It showed that the gas velocity and catalyst loading were two crucial operating parameters to control the process. The results reported here can provide guidance for the operation and reactor design of the CO2 photoreduction process.


Quality assurance, benchmarking, assessment and mutual international recognition of qualifications.
The aim of this report is to provide guidance to assist in the international convergence of quality assurance, benchmarking and assessment systems to improve dental education. Proposals are developed for mutual recognition of qualifications, to aid international movement and exchange of staff and students including and supporting developing countries. Quality assurance is the responsibility of all staff involved in dental education and involves three levels: internal, institutional and external. Benchmarking information provides a subject framework. Benchmarks are useful for a variety of purposes including design and validation of programmes, examination and review; they can also strengthen the accreditation process undertaken by professional and statutory bodies. Benchmark information can be used by institutions as part of their programme approval process, to set degree standards. The standards should be developed by the dental academic community through formal groups of experts. Assessment outcomes of student learning are a measure of the quality of the learning programme. The goal of an effective assessment strategy should be that it provides the starting point for students to adopt a positive approach to effective and competent practice, reflective and lifelong learning. All assessment methods should be evidence based or based upon research. Mutual recognition of professional qualifications means that qualifications gained in one country (the home country) are recognized in another country (the host country). It empowers movement of skilled workers, which can help resolve skills shortages within participating countries. These proposals are not intended to be either exhaustive or prescriptive; they are purely for guidance and derived from the identification of what is perceived to be 'best practice'.


Optimising care for patients with cognitive impairment and dementia following hip fracture.
The global shift in demographics towards aging populations is leading to a commensurate increase in age-related disease and frailty. It is essential to optimise health services to meet current needs and prepare for anticipated future demands. This paper explores issues impacting on people living with cognitive impairment and/or dementia who experience a hip fracture and are cared for in acute settings. This is important given the high mortality and morbidity associated with this population. Given the current insufficiency of clear evidence on optimum rehabilitation of this patient group, this paper explored three key themes namely: recognition of cognitive impairment, response by way of training and education of staff to optimise care for this patient group and review of the importance of outcomes measures. Whilst there is currently insufficient evidence to draw conclusions about the optimal ways of caring for patients living with dementia following hip fracture, this paper concludes that future research should improve understanding of healthcare staff education to improve the outcomes for this important group of patients.


Audit of referral practice to a fast-access breast clinic before the guaranteed 2-week wait.
Currently in the fast-access breast clinic at Eastbourne District General Hospital, the specialist determines the level of urgency of a referral. With the new '2-week wait' imposed by the Government since 1 April 1999, the determination of urgency has transferred to the general practitioner. Therefore, we decided to audit the current situation to see whether the guidelines were adequate for this change of emphasis. A total of 100 consecutive patients referred to the fast-access breast clinic were evaluated to assess the quality of referrals and the effectiveness of the clinic. Only 80% of the referrals adhered to the guidelines; 73% of the referrals were deemed by the specialist to be urgent and seen within the 2-week time period. All patients who were subsequently diagnosed as having breast cancer were seen within 2 weeks. The audit indicates that further specific changes could be made to the guidelines to improve the referral practice.


Association of the gene polymorphisms IFN-gamma +874, IL-13 -1055 and IL-4 -590 with patterns of reinfection with Schistosoma mansoni.
The immunologic findings that most consistently correlate with resistance in human schistosomiasis are high levels of IgE and low levels of IgG4. We have genotyped gene and promoter polymorphisms of cytokines associated with regulation of these isotypes in a cohort of men occupationally exposed to Schistosoma mansoni in western Kenya and evaluated their patterns with respect to resistance and susceptibility to reinfection after treatment and cure with praziquantel (PZQ). In this cohort, polymorphisms in IL-4 (-590T high IgE), IL-13 (-1055T high producer) and IFN-gamma (+874A high producer) demonstrated several correlations with resistance to reinfection. Resistance to reinfection was significantly correlated with the heterozygous IL-4 -590 genotype C/T (OR 3.5, [CI 1.2, 10.2]) compared to T/T. Among men with a homozygous IL-13 genotype CC/TT, having a T allele at the IFN-gamma +874 position increased the odds of resistance relative to individuals with the IFN-gamma +874 A/A genotype (OR = 17.5 [CI 3.0, 101.5]). Among men with homozygous A/A IFN-gamma genotype, the heterozygous IL-13 genotype C/T was associated with resistance relative to the homozygous C/C or T/T genotypes (OR = 22.5 [CI 3.5, 144.4]). No increases in odds of resistance were found in relation to the IL-13 genotype among those with a T allele in the IFN-gamma gene or in relation to the IFN-gamma genotype among those with a heterozygous IL-13 genotype. Calculation of the attributable proportion of resistance showed a significant synergistic interaction between IL-13 -1055 C/T and IL-4 -590 C/T. The identified polymorphisms do not by themselves confer resistance or susceptibility, but we propose that these genotypes allow the resistant phenotype to be developed and expressed upon suitable immune exposure. Based on the literature, these polymorphisms contribute to the regulation of their respective cytokines, likely leading to downstream differences in the production and interrelationships of critical defense mechanisms.


Macrocephaly as a clinical indicator of genetic subtypes in autism.
An association between autism and macrocephaly has been previously described. A subset of cases with extreme macrocephaly (>3 standard deviation [SD], 99.7th percentile) have been correlated to mutations in the gene phosphatase and tensin homolog (PTEN). However, the phenotypic and genetic characterization of the remaining cases remains unclear. We report the phenotypic classification and genetic testing evaluation of a cohort of 33 patients with autism and macrocephaly. Within our cohort, we confirm the association of PTEN mutations and extreme macrocephaly (>3 SD, 99.7th percentile) and identify mutations in 22% of cases, including three novel PTEN mutations. In addition, we define three phenotypic subgroups: (a) those cases associated with somatic overgrowth, (b) those with disproportionate macrocephaly, and (c) those with relative macrocephaly. We have devised a novel way to segregate patients into these subgroups that will aide in the stratification of autism macrocephaly cases. Within these subgroups, we further expand the genetic etiologies for autism cases with macrocephaly by describing two novel suspected pathogenic copy number variants located at 6q23.2 and 10q24.32. These findings demonstrate the phenotypic heterogeneity of autism cases associated with macrocephaly and their genetic etiologies. The clinical yield from PTEN mutation analysis is 22% and 9% from chromosomal microarray (CMA) testing within this cohort. The identification of three distinct phenotypic subgroups within macrocephaly autism patients may allow for the identification of their respective distinct genetic etiologies that to date have remained elusive.


Recombinant scorpine: a multifunctional antimicrobial peptide with activity against different pathogens.
Scorpine is an antimicrobial peptide whose structure resembles a hybrid between a defensin and a cecropin. It exhibits antibacterial activity and inhibits the sporogonic development of parasites responsible for murine malaria. In this communication we report the production of scorpine in a heterelogous system, using a specific vector containing its cloned gene. The recombinantly expressed scorpine (RScp) in (Anopheles gambie) cells showed antibacterial activity against (Bacillus subtilis) and (Klebsiella pneumoniae), at 5 and 10 microM, respectively. It also produced 98% mortality in sexual stages of (Plasmodium berghei) at 15 microM and 100% reduction in (Plasmodium falciparum) parasitemia at 5 microM. RScp also inhibited virus dengue-2 replication in C6/36 mosquito cells. In addition, we generated viable and fertile transgenic (Drosophila) that overexpresses and correctly secretes RScp into the insect hemolymph, suggesting that the generation of transgenic mosquitoes resistant to different pathogens may be viable.


Using ultrasound to guide the insertion of pedicle screws during scoliosis surgery.
Scoliosis surgery involves the insertion of screws and/or hooks into selected vertebrae to secure a pre-bent rod placed along the concave side of the spine. Usually conventional x-rays will be taken before the surgery to plan the alignment and positioning of the pedicle screws. However, reports state that perforation rate range from 6% to 54%. A misalignment of a pedicle screw can potentially cause permanent neurological spinal cord injury or even a life-threatening vascular injury. Because of the importance of positioning and aligning of pedicle screws, we are working on an ultrasound method to guide the insertion of pedicle screws in real time. A pulse-echo immersion experiment was set up to study how well the edges of cortical bone could be detected using a bovine spinous process in-vitro. Two ultrasound frequencies (3.5 MHz and 5.0 MHz) were considered in this study. This preliminary study shows that ultrasound is able to penetrate cortical bone and reflect back from the outer boundary. All interfaces are clearly identified for both frequencies. Strong reflection signals are obtained when the beam is normal to the interface. Derived thickness values from the reflections are comparable with those from micro-CT image. The 5.0 MHz ultrasound frequency provided better resolution than the 3.0 MHz frequency.


Systematic Screening of Promoter Regions Pinpoints Functional Cis-Regulatory Mutations in a Cutaneous Melanoma Genome.
With the recent discovery of recurrent mutations in the TERT promoter in melanoma, identification of other somatic causal promoter mutations is of considerable interest. Yet, the impact of sequence variation on the regulatory potential of gene promoters has not been systematically evaluated. This study assesses the impact of promoter mutations on promoter activity in the whole-genome sequenced malignant melanoma cell line COLO-829. Combining somatic mutation calls from COLO-829 with genome-wide chromatin accessibility and histone modification data revealed mutations within promoter elements. Interestingly, a high number of potential promoter mutations (n = 23) were found, a result mirrored in subsequent analysis of TCGA whole-melanoma genomes. The impact of wild-type and mutant promoter sequences were evaluated by subcloning into luciferase reporter vectors and testing their transcriptional activity in COLO-829 cells. Of the 23 promoter regions tested, four mutations significantly altered reporter activity relative to wild-type sequences. These data were then subjected to multiple computational algorithms that score the cis-regulatory altering potential of mutations. These analyses identified one mutation, located within the promoter region of NDUFB9, which encodes the mitochondrial NADH dehydrogenase (ubiquinone) 1 beta subcomplex 9, to be recurrent in 4.4% (19 of 432) of TCGA whole-melanoma exomes. The mutation is predicted to disrupt a highly conserved SP1/KLF transcription factor binding motif and its frequent co-occurrence with mutations in the coding sequence of NF1 supports a pathologic role for this mutation in melanoma. Taken together, these data show the relatively high prevalence of promoter mutations in the COLO-829 melanoma genome, and indicate that a proportion of these significantly alter the regulatory potential of gene promoters. Genomic-based screening within gene promoter regions suggests that functional cis-regulatory mutations may be common in melanoma genomes, highlighting the need to examine their role in tumorigenesis.


Vaccine hesitancy among healthcare workers in Europe: A qualitative study.
Healthcare workers (HCWs) are often referred to as the most trusted source of vaccine-related information for their patients. However, the evidence suggests that a number of HCWs are vaccine-hesitant. This study consists of 65 semi-structured interviews with vaccine providers in Croatia, France, Greece, and Romania to investigate concerns HCWs might have about vaccination. The results revealed that vaccine hesitancy is present in all four countries among vaccine providers. The most important concern across all countries was the fear of vaccine side effects. New vaccines were singled out due to perceived lack of testing for vaccine safety and efficacy. Furthermore, while high trust in health authorities was expressed by HCWs, there was also strong mistrust of pharmaceutical companies due to perceived financial interests and lack of communication about side effects. The notion that it is a doctor's responsibility to respond to hesitant patients was reported in all countries. Concerns were also seen to be country- and context-specific. Strategies to improve confidence in vaccines should be adapted to the specific political, social, cultural and economic context of countries. Furthermore, while most interventions focus on education and improving information about vaccine safety, effectiveness, or the need for vaccines, concerns raised in this study identify other determinants of hesitancy that need addressing. The representativeness of the views of the interviewed HCWs must be interpreted with caution. This a qualitative study with a small sample size that included geographical areas where vaccination uptake was lower or where hesitancy was more prevalent and it reflects individual participants' beliefs and attitudes toward the topic. As HCWs have the potential of influencing patient vaccination uptake, it is crucial to improve their confidence in vaccination and engage them in activities targeting vaccine hesitancy among their patients.


Functions of the water soluble chlorophyll-binding protein in plants.
Functional aspects of water soluble chlorophyll-binding protein (WSCP) in plants were investigated during the courses of leaf senescence, chlorophyll biogenesis, stress response and photoprotection. The cDNA sequence encoding WSCP from cauliflower was cloned into a binary vector to facilitate Agrobacterium tumefaciens mediated transformation of Nicotiana tabacum. The resultant transgenic tobacco plants overexpressed the CauWSCP gene under the control of a 35S-promoter. Analyses of protein and pigment contents indicate that WSCP overexpression does not enhance chlorophyll catabolism in vivo, thus rendering a role of WSCP in Chl degradation unlikely. Accumulation of higher levels of protochlorophyllide in WSCP overexpressor plants corroborates a proposed temporary storage and carrier function of WSCP for chlorophyll and late precursors. Although WSCP overexpressor plants did not show significant differences in non-photochemical quenching of chlorophyll fluorescence, they are characterized by significantly lower zeaxanthin accumulation and peroxidase activity at different light intensities, even at high light intensities of 700-900μmol photons m(-2)s(-1). These results suggest a photoprotective function of the functional chlorophyll binding-WSCP tetramer by shielding of chlorophylls from molecular oxygen.


Analysis of the Diffusivity Change from Single-Molecule Trajectories on Living Cells.
With the wide application of live-cell single-molecule imaging and tracking of biomolecules at work, deriving diffusion state changes of individual molecules is of particular interest as these changes reflect molecular oligomerization or interaction with other cellular components and thus correlate with functional changes. We have developed a Rayleigh mixture distribution-based hidden Markov model method to analyze time-lapse diffusivity change of single molecules, especially membrane proteins, with unknown dynamic states in living cells. With this method, the diffusion parameters, including diffusion state number, state transition probability, diffusion coefficient, and state mixture ratio, can be extracted from the single-molecule diffusion trajectories accurately via easy computation. The validity of our method has been demonstrated with not only experiments on synthetic trajectories but also single-molecule fluorescence imaging data of two typical membrane receptors. Our method offers a new analytical tool for the investigation of molecular interaction kinetics at the single-molecule level.


CMAUP database update 2024: extended functional and association information of useful plants for biomedical research.
Knowledge of the collective activities of individual plants together with the derived clinical effects and targeted disease associations is useful for plant-based biomedical research. To provide the information in complement to the established databases, we introduced a major update of CMAUP database, previously featured in NAR. This update includes (i) human transcriptomic changes overlapping with 1152 targets of 5765 individual plants, covering 74 diseases from 20 027 patient samples; (ii) clinical information for 185 individual plants in 691 clinical trials; (iii) drug development information for 4694 drug-producing plants with metabolites developed into approved or clinical trial drugs; (iv) plant and human disease associations (428 737 associations by target, 220 935 reversion of transcriptomic changes, 764 and 154121 associations by clinical trials of individual plants and plant ingredients); (v) the location of individual plants in the phylogenetic tree for navigating taxonomic neighbors, (vi) DNA barcodes of 3949 plants, (vii) predicted human oral bioavailability of plant ingredients by the established SwissADME and HobPre algorithm, (viii) 21-107% increase of CMAUP data over the previous version to cover 60 222 chemical ingredients, 7865 plants, 758 targets, 1399 diseases, 238 KEGG human pathways, 3013 gene ontologies and 1203 disease ontologies. CMAUP update version is freely accessible at https://bidd.group/CMAUP/index.html.


To Be or Not to Be Expressed: The First Evidence of a Nucleolar Dominance Tissue-Specificity in Brachypodium hybridum.
Nucleolar dominance (ND) is an epigenetic, developmentally regulated phenomenon that describes the selective inactivation of 35S rDNA loci derived from one progenitor of a hybrid or allopolyploid. The presence of ND was documented in an allotetraploid grass, Brachypodium hybridum (genome composition DDSS), which is a polyphyletic species that arose from crosses between two putative ancestors that resembled the modern B. distachyon (DD) and B. stacei (SS). In this work, we investigated the developmental stability of ND in B. hybridum genotype 3-7-2 and compared it with the reference genotype ABR113. We addressed the question of whether the ND is established in generative tissues such as pollen mother cells (PMC). We examined condensation of rDNA chromatin by fluorescence in situ hybridization employing state-of-art confocal microscopy. The transcription of rDNA homeologs was determined by reverse-transcription cleaved amplified polymorphic sequence analysis. In ABR113, the ND was stable in all tissues analyzed (primary and adventitious root, leaf, and spikes). In contrast, the 3-7-2 individuals showed a strong upregulation of the S-genome units in adventitious roots but not in other tissues. Microscopic analysis of the 3-7-2 PMCs revealed extensive decondensation of the D-genome loci and their association with the nucleolus in meiosis. As opposed, the S-genome loci were always highly condensed and localized outside the nucleolus. These results indicate that genotype-specific loss of ND in B. hybridum occurs probably after fertilization during developmental processes. This finding supports our view that B. hybridum is an attractive model to study ND in grasses.


Perforin, a cytotoxic molecule which mediates cell necrosis, is not required for the early control of mycobacterial infection in mice.
Host defense against mycobacterial infection requires the participation of monocytes and T cells. Both CD4+ and CD8+ T cells have been shown to be important in resistance to mycobacterial infection in vivo. The main contribution of CD4+ T cells to the protective antituberculosis response involves the production of Th1-type cytokines, including interleukin-2 (IL-2) and gamma interferon (IFN-gamma). CD8+ T cells have been considered to be responsible primarily for cytotoxicity mediated by toxic molecules, including perforin. CD8+ T cells may also elaborate Th1-type cytokines, such as IFN-gamma, in response to the infection. To elucidate the contribution of perforin-mediated target cell death to the control of mycobacterial infection in vivo, mice with a disruption in the perforin gene (P-/-) were infected with Mycobacterium bovis BCG or M. tuberculosis Erdman for 5 and 13 weeks, respectively. At 1, 3, 5, and 13 weeks postinfection, the number of viable mycobacteria in the lungs, spleens, and livers of mice were determined by CFU assay. The infected tissues were examined histologically, and cytokine mRNA levels in the spleens of these mice were determined. Similar studies were carried out in Fas receptor-defective (CBA/lpr(cg)) mice to evaluate the contribution of this alternative cytotoxic pathway to the control of mycobacterial infection. The absence of either perforin gene function or Fas receptor gene function did not modify the course of experimental mycobacterial infection in these mice. In addition, both P-/- and Fas receptor-defective mice appeared to have a compensatory activation of cytokine genes, even in the absence of the experimental infection. P-/- mice had a mean 3.4- to 5-fold increase in mRNA levels for IL-10, IL-12p35, IL-6, and IFN-gamma. Similarly, Fas receptor-defective mice had a mean 3- to 3.6-fold increase in mRNA levels for IFN-gamma, IL-12p35, and IL-10. Our results indicate that both perforin-mediated cytotoxicity and Fas-mediated cytotoxicity do not appear to be necessary for the early control of mycobacterial infection in vivo.


Generation of an induced pluripotent stem cell line (CSC-46) from a patient with Parkinson's disease carrying a novel p.R301C mutation in the GBA gene.
Mutations in the glucocerebrosidase (GBA) gene have been associated with the development of Parkinson's disease (PD). An induced pluripotent stem cell (iPSC) line was generated from a 60-year old patient diagnosed with PD and carrying a new mutation variant p.R301C in GBA. Using non-integrating Sendai virus-based technology, we utilized OCT3/4, SOX2, c-MYC and KLF4 transcription factors to reprogram skin fibroblasts into iPSCs. The generated iPSC line retained the mutation, displayed expression of common pluripotency markers, differentiated into the three germ layers, and exhibited normal karyotype. The iPSC line can be further used for studying PD pathogenesis.


Improving comfort around dying in elderly people: a cluster randomised controlled trial.
Over 50% of elderly people die in acute hospital settings, where the quality of end-of-life care is often suboptimum. We aimed to assess the effectiveness of the Care Programme for the Last Days of Life (CAREFuL) at improving comfort and quality of care in the dying phase in elderly people. We did a cluster randomised controlled trial in acute geriatric wards in ten hospitals in Flemish Region, Belgium, between Oct 1, 2012, and March 31, 2015. Hospitals were randomly assigned to implementation of CAREFuL (CAREFuL group) or to standard care (control group) using a random number generator. Patients and families were masked to interventaion allocation; hospital staff were unmasked. CAREFuL comprised a care guide for the last days of life, training, supportive documentation, and an implementation guide. Primary outcomes were comfort around dying, measured with the End-of-Life in Dementia-Comfort Assessment in Dying (CAD-EOLD), and symptom management, measured with the End-of-Life in Dementia-Symptom Management (SM-EOLD), by nurses and family carers. Analyses were by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT01890239. 451 (11%) of 4241 beds in ten hospitals were included in the analyses. Five hospitals were randomly assigned to standard health care practice and five to the CAREFuL programme; 118 patients in the control group and 164 in the CAREFuL group were eligible for assessment. Assessments were done for 132 (80%) of 164 patients in the CAREFuL group and 109 (92%) of 118 in the control group by nurses, and 48 (29%) in the CAREFuL group and 23 (19%) in the control group by family carers. Implementation of CAREFuL compared with control significantly improved nurse-assessed comfort (CAD-EOLD baseline-adjusted mean difference 4·30, 95% CI 2·07-6·53; p<0·0001). No significant differences were noted for the CAD-EOLD assessed by family carers (baseline-adjusted mean difference -0·62, 95% CI -6·07 to 4·82; p=0·82) or the SM-EOLD assessed by nurses (-0·41, -1·86 to 1·05; p=0·58) or by family carers (-0·59, -3·75 to 2·57; p=0·71). Although a continuous monitoring of the programme is warranted, these results suggest that implementation of CAREFuL might improve care during the last days of life for patients in acute geriatric hospital wards. The Flemish Government Agency for Innovation by Science and Technology and the Belgian Cancer Society "Kom Op Tegen Kanker".


Transmission of wild-type and recombinant HaSNPV among larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) on cotton.
Horizontal transmission of insect viruses is a key factor in their cycling in agro-ecosystems. Here we study the transmission of the baculovirus HaSNPV among larvae of Helicoverpa armigera (Hübner) in cotton. Transmission of three HaSNPV genotypes was studied from larvae infected with a single virus genotype and from larvae infected with two different genotypes. Genotypes included a wild-type virus, an ecdysteroid UDP-glucosyltransferase (egt) deletion mutant (HaSNPV-LM2) with slightly enhanced speed of kill, and an egt-negative genotype that expresses a neurotoxin gene derived from the scorpion Androctonus australis Hector (HaSNPV-4A). The latter genotype has a substantially increased speed of kill. In three field experiments, the wild-type and egt deletion virus variants and a mixture of the two had similar rates of transmission. Transmission increased with density of infector insects and decreased with time lapsed since the inoculation of the infector larvae. Transmission of the neurotoxin expressing virus was lower than that of the other two genotypes in a glasshouse experiment. The studied genotypes of HaSNPV have significant differences in time to kill and virus yield, but we found no significant differences in rates of virus transmission at the crop level in the case of the egt deletion variant HaSNPV-LM2. Transmission of the transgenic virus genotype HaSNPV-4A was significantly reduced. Overall, differences in transmission between virus genotypes were subtler, and more difficult to detect with statistical significance, than effects of other factors, such as density of infectors and time delay between release of infectors and recipient caterpillars on the plant.


Viral evolution and interferon resistance of hepatitis C virus RNA replication in a cell culture model.
Hepatitis C virus (HCV) replicates through an error-prone process that may support the evolution of genetic variants resistant to the host cell antiviral response and interferon (IFN)-based therapy. We evaluated HCV-IFN interactions within a long-term culture system of Huh7 cell lines harboring different variants of an HCV type 1b subgenomic RNA replicon that differed at only two sites within the NS5A-encoding region. A replicon with a K insertion at HCV codon 2040 replicated efficiently and exhibited sequence stability in the absence of host antiviral pressure. In contrast, a replicon with an L2198S point mutation replicated poorly and triggered a cellular response characterized by IFN-beta production and low-level IFN-stimulated gene (ISG) expression. When maintained in long term-culture, the L2198S RNA evolved into a stable high-passage (HP) variant with six additional point mutations throughout the HCV protein-encoding region that enhanced viral replication. The HP RNA transduced Huh7 cells with more than 1,000-fold greater efficiency than its L2198S progenitor or the K2040 sequence. Replication of the HP RNA resisted suppression by IFN-alpha treatment and was associated with virus-directed reduction in host cell expression of ISG56, an antagonist of HCV RNA translation. Accordingly, the HP RNA was retained within polyribosome complexes in vivo that were refractory to IFN-induced disassembly. These results identify ISG56 as a translational control effector of the host response to HCV and provide direct evidence to link this response to viral sequence evolution, ISG regulation, and selection of the IFN-resistant viral phenotype.


[Anesthesiologic peculiarities in bilateral pheochromocytoma and suspected multiple endocrine neoplasia].
Based on a case report with surgical removal of bilateral giant phaeochromocytomas with heredofamilial affliction and suspicion of multiple endocrine neoplasia type II (MEN-II syndrome) and of malignancy, the anaesthetic implications are discussed in a patient inadvertently not prepared by antiadrenergic treatment. It is pointed out that in MEN II syndrome, a combination of phaeochromocytoma and medullary carcinoma of the thyroid, surgical removal of the phaeochromocytoma merits utmost priority before thyreoidectomy. The most important laboratory parameters in the diagnosis of MEN II syndrome are pre- and postoperative determinations of calcitonin and carcinoembryonal antigen (CEA).