A Culture Collection of 50 Neisseria gonorrhoeae Isolates.

A culture collection of 50 Neisseria gonorrhoeae isolates is available from the CDC & FDA Antibiotic Resistance Isolate Bank. Associated data include antibiotic susceptibility information for azithromycin, cefixime, cefpodoxime, ceftriaxone, tetracycline, ciprofloxacin, penicillin, and spectinomycin and linked whole-genome sequences.

Adherence to Guideline Recommendations for Perioperative Chemotherapy in Patients with pN2-3 M0 Squamous Cell Carcinoma of the Penis: Temporal Trends and Survival Outcomes.

AIMS: To analyse contemporary perioperative chemotherapy (CHT) guideline adherence rates for pN2-3 M0 squamous cell carcinoma of the penis, as well as CHT association with cancer-specific (CSM) and other-cause mortality (OCM). MATERIALS AND METHODS: Within the Surveillance, Epidemiology, and End Results databases, 311 pN2-3 M0 squamous cell carcinoma of the penis patients treated with inguinal lymph node dissection were identified. Univariable and multivariable logistic regression analyses focused on CHT rates, whereas cumulative incidence plots and multivariable competing risks regression analyses tested for CSM and OCM rates. RESULTS: CHT was administered to 140 (45%) patients and rates increased from 37.5 to 62.2% (2004-2015; P = 0.02). Specifically, annual CHT rates increased over time in patients younger or equal to 65 years and in patients older than 65 years (44.4-84.6% versus 28.6-50%, respectively), but this trend was not statistically significant (P = 0.1 and P = 0.2, respectively). The median follow-up was 13 months for both CHT (interquartile range 8.0-32.2) and no-CHT subgroups (interquartile range 5.0-40.0). In multivariable logistic regression analyses, more contemporary year of diagnosis interval (odds ratio 2.08, P < 0.01) and age older than 75 years (odds ratio 0.14, P < 0.001) were independent predictors of CHT use. In multivariable competing risks regression analyses, CHT use did not affect CSM (hazard ratio 1.02; P = 0.7) or OCM (hazard ratio 1.56; P = 0.8). CONCLUSIONS: CHT adherence rates sharply increased in the most recent years. Despite this increase over time, the lack of efficacy regarding CSM benefit is disappointing.

Maternal high-fat diet alters anxiety behavior and glucocorticoid signaling in adolescent offspring.

Maternal obesity and overconsumption of saturated fats during pregnancy have profound effects on offspring health, ranging from metabolic to behavioral disorders in later life. The influence of high-fat diet (HFD) exposure on the development of brain regions implicated in anxiety behavior is not well understood. We previously found that maternal HFD exposure is associated with an increase in anxiety behavior and alterations in the expression of several genes involved in inflammation via the glucocorticoid signaling pathway in adult rat offspring. During adolescence, the maturation of feedback systems mediating corticosteroid sensitivity is incomplete, and therefore distinct from adulthood. In this study, we examined the influence of maternal HFD on several measures of anxiety behavior and gene expression in adolescent offspring. We examined the expression of corticosteroid receptors and related inflammatory processes, as corticosteroid receptors are known to regulate circulating corticosterone levels during basal and stress conditions in addition to influencing inflammatory processes in the hippocampus and amygdala. We found that adolescent animals perinatally exposed to HFD generally showed decreased anxiety behavior accompanied by a selective alteration in the expression of the glucocorticoid receptor and several downstream inflammatory genes in the hippocampus and amygdala. These data suggest that adolescence constitutes an additional period when the effects of developmental programming may modify mental health trajectories.

Exercise and environment as an intervention for neonatal alcohol effects on hippocampal adult neurogenesis and learning.

Neonatal alcohol exposure impairs cognition and learning in adulthood and permanently damages the hippocampus. Wheel running (WR) improves hippocampus-associated learning and memory and increases the genesis and survival of newly generated neurons in the hippocampal dentate gyrus. WR significantly increases proliferation of newly generated dentate granule cells in alcohol-exposed (AE) and control rats on Postnatal Day (PD) 42 but only control rats show an increased number of surviving cells thirty days after WR (Helfer et al., 2009b). The present studies examined whether proliferation-promoting WR followed by survival-enhancing environmental complexity (EC) during adolescence could increase survival of new neurons in AE rats. On PD 4-9, pups were intubated with alcohol in a binge-like manner (5.25g/kg/day, AE), were sham-intubated (SI), or were reared normally (suckle control, SC). On PD 30 animals were assigned to WR (PD 30-42) followed by EC (PD 42-72; WR/EC) or were socially housed (SH/SH) for the duration of the experiment. All animals were injected with 200mg/kg bromodeoxyuridine (BrdU) on PD 41. In Experiment 1, survival of newly generated cells was significantly enhanced in the AE-WR/EC group in comparison with AE-SH/SH group. Experiment 2A examined trace eyeblink conditioning. In the SH/SH condition, AE impaired trace eyeblink conditioning relative to SI and SC controls. In the WR/EC condition, AE rats performed as well as controls. In Experiment 2B, the same intervention was examined using the context preexposure facilitation effect (CPFE); a hippocampus-dependent variant of contextual fear conditioning. Again, the WR/EC intervention reversed the deficit in conditioned fear to the context that was evident in the SH/SH condition. Post-weaning environmental manipulations promote cell survival and reverse learning deficits in rats that were exposed to alcohol during development. These manipulations may provide a basis for developing interventions that ameliorate learning impairments associated with human fetal alcohol spectrum disorders.

Comparison of success rates, learning curves, and inter-subject performance variability of robot-assisted and manual ultrasound-guided nerve block needle guidance in simulation.

BACKGROUND: This study focuses on a recently developed robotic nerve block system and its impact on learning regional anaesthesia skills. We compared success rates, learning curves, performance times, and inter-subject performance variability of robot-assisted vs manual ultrasound (US)-guided nerve block needle guidance. The hypothesis of this study is that robot assistance will result in faster skill acquisition than manual needle guidance. METHODS: Five co-authors with different experience with nerve blocks and the robotic system performed both manual and robot-assisted, US-guided nerve blocks on two different nerves of a nerve phantom. Ten trials were performed for each of the four procedures. Time taken to move from a shared starting position till the needle was inserted into the target nerve was defined as the performance time. A successful block was defined as the insertion of the needle into the target nerve. Average performance times were compared using analysis of variance. P<0.05 was considered significant. Data presented as mean (standard deviation). RESULTS: All blocks were successful. There were significant differences in performance times between co-authors to perform the manual blocks, either superficial (P=0.001) or profound (P=0.0001); no statistical difference between co-authors was noted for the robot-assisted blocks. Linear regression indicated that the average decrease in time between consecutive trials for robot-assisted blocks of 1.8 (1.6) s was significantly (P=0.007) greater than the decrease for manual blocks of 0.3 (0.3) s. CONCLUSIONS: Robot assistance of nerve blocks allows for faster learning of needle guidance over manual positioning and reduces inter-subject performance variability.

Perinatal high fat diet alters glucocorticoid signaling and anxiety behavior in adulthood.

Maternal obesity carries significant health risks for offspring that manifest later in life, including metabolic syndrome, cardiovascular disease and affective disorders. Programming of the hypothalamic-pituitary-adrenal (HPA) axis during development mediates both metabolic homeostasis and the response to psychosocial stress in offspring. A diet high in fat alters maternal systemic corticosterone levels, but effects in offspring on limbic brain areas regulating the HPA axis and anxiety behavior are poorly understood. In addition to their role in the response to psychosocial stress, corticosteroid receptors form part of the glucocorticoid signaling pathway comprising downstream inflammatory processes. Increased systemic inflammation is a hallmark of high-fat diet exposure, though altered expression of these genes in limbic brain areas has not been examined. We studied the influence of high-fat diet exposure during pre-weaning development in rats on gene expression in the amygdala and hippocampus by quantitative real-time polymerase chain reaction (PCR), anxiety behavior in the Open field, elevated plus maze and light-dark transition tasks, and corticosterone levels in response to stress by radioimmunoassay. As adults, offspring exposed to perinatal high-fat diet show increased expression of corticosterone receptors in the amygdala and altered pro-inflammatory and anti-inflammatory expression in the hippocampus and amygdala in genes known to be regulated by the glucocorticoid receptor. These changes were associated with increased anxiety behavior, decreased basal corticosterone levels and a slower return to baseline levels following a stress challenge. The data indicate that the dietary environment during development programs glucocorticoid signaling pathways in limbic areas relevant for the regulation of HPA function and anxiety behavior.

Transcontinental anaesthesia: a pilot study.

BACKGROUND: Although telemedicine is one of the key initiatives of the World Health Organization, no study has explored the feasibility and efficacy of teleanaesthesia. This bi-centre pilot study investigates the feasibility of transcontinental anaesthesia. METHODS: Twenty patients aged ≥ 18 yr undergoing elective thyroid surgery for ≥ 30 min were enrolled in this study. The remote and local set-up was composed of a master-computer (Montreal) and a slave-computer (Pisa). Standard Internet connection, remote desktop control, and video conference software were used. All patients received total i.v. anaesthesia controlled remotely (Montreal). The main outcomes were feasibility, clinical performance, and controller performance of transcontinental anaesthesia. The clinical performance of hypnosis control was the efficacy to maintain bispectral index (BIS) at 45: 'excellent', 'good', 'poor', and 'inadequate' control represented BIS values within 10, from 11 to 20, from 21 to 30, or >30% from target. The clinical performance of analgesia was the efficacy to maintain Analgoscore values at 0 (-9 to 9); -3 to +3 representing 'excellent' pain control, -3 to -6 and +3 to +6 representing 'good' pain control, and -6 to -9 and +6 to +9 representing 'insufficient' pain control. The controller performance was evaluated using Varvel parameters. RESULTS: Transcontinental anaesthesia was successful in all 20 consecutive patients. The clinical performance of hypnosis showed an 'excellent and good' control for 69% of maintenance time, and the controller performance showed an average global performance index of 57. The clinical performance of analgesia was 'excellent and good' for 92% of maintenance time, and the controller performance showed a global performance index of 1118. CONCLUSIONS: Transcontinental anaesthesia is feasible; control of anaesthesia shows good performance indexes. Clinical registration number NCT01331096.

Evaluation of a novel closed-loop total intravenous anaesthesia drug delivery system: a randomized controlled trial.

BACKGROUND: We have developed an automatic anaesthesia system for closed-loop administration of anaesthesia drugs. The control variables used were bispectral index (BIS) and Analgoscore for hypnosis and antinociception, respectively. METHODS: One hundred and eighty-six patients were randomly enrolled in two groups. Propofol, remifentanil, and rocuronium were administered using closed-loop feedback control (closed-loop, n = 93) or manually (control group, n = 93). The clinical performance of hypnosis control was determined by calculating the offset from a BIS of 45: 'excellent', 'good', 'poor', and 'inadequate' control was defined as BIS values within 10%, from 11% to 20%, from 21% to 30%, or >30% offset from the target. The clinical performance of analgesia was defined as the offset from Analgoscore values. Data presented as mean (standard deviation) (95% confidence interval). RESULTS: Excellent or good control of hypnosis was achieved significantly longer in the closed-loop group [47.0 (9.8%) (45.0/49.0), 34.4 (4.7%) (33.5/35.4)] than in the control group [37.3 (14.3%) (34.4/40.2) and 32.3 (7.6%) (30.7/33.7)], respectively (P<0.0001 and 0.0085). Poor and inadequate control of hypnosis was significantly shorter in the closed-loop group [10.8 (5.0%) (9.8/11.8) and 7.7 (6.2%) (6.4/9.0)] than in the control group [14.7 (6.8%) (13.3/16.0) and 15.8 (14.7%) (12.8/18.8)], respectively (P<0.0001). Excellent control of analgesia was achieved significantly longer in the closed-loop group [78.7 (16.2%) (75.4/82.0)] than in the control group [73.7 (17.8%) (70.1/77.3)] (P=0.0456). CONCLUSIONS: The closed-loop system was better at maintaining BIS and Analgoscore than manual administration.

Effects of exercise and environmental complexity on deficits in trace and contextual fear conditioning produced by neonatal alcohol exposure in rats.

In rodents, voluntary exercise and environmental complexity increases hippocampal neurogenesis and reverses spatial learning and long-term potentiation deficits in animals prenatally exposed to alcohol. The present experiment extended these findings to neonatal alcohol exposure and to delay, trace, and contextual fear conditioning. Rats were administered either 5.25 g/kg/day alcohol via gastric intubation or received sham-intubations (SI) between Postnatal Day (PD) 4 and 9 followed by either free access to a running wheel on PD 30-41 and housing in a complex environment on PD 42-72 (wheel-running plus environmental complexity; WREC) or conventional social housing (SHSH) from PD 30 to 72. Adult rats (PD 80 ± 5) received 5 trials/day of a 10-s flashing-light conditioned stimulus (CS) paired with .8 mA footshock either immediately (delay conditioning) or after a 10-s trace interval (trace conditioning) for 2 days. Neonatal alcohol exposure impaired context and trace conditioning, but not short-delay conditioning. The WREC intervention did not reverse these deficits, despite increasing context-related freezing in ethanol-exposed and SI animals.

First robotic tracheal intubations in humans using the Kepler intubation system.

BACKGROUND: Intubation is one of the most important anaesthetic skills. We developed a robotic intubation system (Kepler intubation system, KIS) for oral tracheal intubation. METHODS: In this pilot study, 12 patients were enrolled after approval of the local Ethics board and written informed consent. The KIS consists of four main components: a ThrustMaster T.Flight Hotas X joystick (Guillemot Inc., New York, NY, USA), a JACO robotic arm (Kinova Rehab, Montreal, QC, Canada), a Pentax AWS video laryngoscope (Ambu A/S, Ballerup, Denmark), and a software control system. The joystick allows simulation of the wrist or arm movements of a human operator. The success rate of intubation and intubation times were measured. RESULTS: Eleven men and one woman aged 66 yr were included in this study. Intubation was successful in all but one patient using KIS at a total time of [median (inter-quartile range; range)] 93 (87, 109; 76, 153) s; in one patient, fogging of the video laryngoscope prevented intubation using KIS. CONCLUSIONS: We present the first human testing of a robotic intubation system for oral tracheal intubation. The success rate was high at 91%. Future studies are needed to assess the performance and safety of such a system.

Reduced cerebral oxygen saturation during thoracic surgery predicts early postoperative cognitive dysfunction.

BACKGROUND: The objective of this prospective study is to determine cognitive dysfunction after thoracic surgery. METHODS: Seventy-six patients undergoing thoracic surgery with single-lung ventilation (SLV) of an expected duration of >45 min were enrolled. Monitoring consisted of standard clinical parameters and absolute oximetry (S(ct)O(2)). The Mini-Mental State Exam (MMSE) test was used to assess cognitive function before operation and at 3 and 24 h after operation. Data were analysed using Spearman correlation test; risks for cognitive dysfunction were expressed as odds ratios. P<0.05 and data are presented as median (interquartile range). RESULTS: One patient was excluded from the study. S(ct)O(2) during SLV decreased to critical values of <65%, 60%, and 55% in 40 (53%), 15 (20%), and 5 patients (7%), respectively. Twenty-two patients (29%) had a decrease of MMSE>2 points 3 h after surgery, eight patients (10%) had a decrease of MMSE>2 points 24 h after surgery. Postoperative cognitive dysfunction correlated at r(2)=0.272, 0.285, 0.297 with patient exposure times to S(ct)O(2)<65% (P=0.018), <60% (P=0.013), <55% (P=0.010), respectively. The odds ratios of developing early cognitive dysfunction ranged from 2.03 (95% CI: 0.74-5.59) for a short (<5 min) exposure to S(ct)O(2)<65% to a maximum of 9.56 (95% CI: 1.75-52.13) when S(ct)O(2) was <60% for more than 30 min. CONCLUSIONS: Early cognitive dysfunction after thoracic surgery with SLV is positively related to intraoperative decline of S(ct)O(2).

Neonatal alcohol exposure disrupts hippocampal neurogenesis and contextual fear conditioning in adult rats.

Developmental alcohol exposure can permanently alter brain structures and produce functional impairments in many aspects of behavior, including learning and memory. This study evaluates the effect of neonatal alcohol exposure on adult neurogenesis in the dentate gyrus of the hippocampus and the implications of such exposure for hippocampus-dependent contextual fear conditioning. Alcohol-exposed rats (AE) received 5.25g/kg/day of alcohol on postnatal days (PD) 4-9 (third trimester in humans), in a binge-like manner. Two control groups were included: sham-intubated (SI) and suckle-control (SC). Animals were housed in social cages (3/cage) after weaning. On PD80, animals were injected with 200mg/kg BrdU. Half of the animals were sacrificed 2h later. The remainder were sacrificed on PD114 to evaluate cell survival; separate AE, SI, and SC rats not injected with BrdU were tested for the context preexposure facilitation effect (CPFE; ~PD117). There was no difference in the number of BrdU+ cells in AE, SI and SC groups on PD80. On PD114, cell survival was significantly decreased in AE rats, demonstrating that developmental alcohol exposure damages new cells' ability to incorporate into the network and survive. Behaviorally tested SC and SI groups preexposed to the training context 24h prior to receiving a 1.5mA 2s footshock froze significantly more during the context test than their counterparts preexposed to an alternate context. AE rats failed to show the CPFE. The current study shows the detrimental, long-lasting effects of developmental alcohol exposure on hippocampal adult neurogenesis and contextual fear conditioning.

Analysis of xylem formation in pine by cDNA sequencing.

Secondary xylem (wood) formation is likely to involve some genes expressed rarely or not at all in herbaceous plants. Moreover, environmental and developmental stimuli influence secondary xylem differentiation, producing morphological and chemical changes in wood. To increase our understanding of xylem formation, and to provide material for comparative analysis of gymnosperm and angiosperm sequences, ESTs were obtained from immature xylem of loblolly pine (Pinus taeda L.). A total of 1,097 single-pass sequences were obtained from 5' ends of cDNAs made from gravistimulated tissue from bent trees. Cluster analysis detected 107 groups of similar sequences, ranging in size from 2 to 20 sequences. A total of 361 sequences fell into these groups, whereas 736 sequences were unique. About 55% of the pine EST sequences show similarity to previously described sequences in public databases. About 10% of the recognized genes encode factors involved in cell wall formation. Sequences similar to cell wall proteins, most known lignin biosynthetic enzymes, and several enzymes of carbohydrate metabolism were found. A number of putative regulatory proteins also are represented. Expression patterns of several of these genes were studied in various tissues and organs of pine. Sequencing novel genes expressed during xylem formation will provide a powerful means of identifying mechanisms controlling this important differentiation pathway.

Molecular cloning and sequence analysis of the penton base genes of type II avian adenoviruses.

We describe here the identification of the penton base gene of hemorrhagic enteritis virus (HEV), a type II avian adenovirus, in a 2477-base pair (bp)-EcoRI fragment of the viral DNA by sequence analysis. Identification is based on an extensive amino acid homology between the HEV-open reading frame and the penton base of a fowl adenovirus (FAV-10) and various human adenoviruses. The 1344 bp-penton base gene of HEV encodes a 448-amino acid polypeptide of molecular weight of 50,843 Da. The nucleotide sequences of penton base genes of HEV and marble spleen disease virus (MSDV) are identical. The HEV penton base lacks the RGD motif, present in most human adenoviruses (Ad2, Ad3, Ad4, and Ad 12) suggesting that HEV may not use alpha v integrins to gain entry into host cells. Further sequence analysis revealed the presence of a Leu-Asp-Val (LDV) motif in the HEV penton base amino acid sequence similar to most of the human adenoviruses. LDV motif on the fibronectin has been shown to interact with the alpha 4 beta 1 integrins on cells, which includes lymphocytes and monocytes. The presence of LDV motif in the penton base of HEV implicates the involvement of alpha 4 beta 1 integrins in the viral internalization into host cells.

Quantitation of human cytomegalovirus glycoprotein H gene in cells using competitive PCR and a rapid fluorescence-based detection system.

A technique is described for quantitation of the human cytomegalovirus (HCMV) glycoprotein H (gH) gene in cells using a quantitative-competitive polymerase chain reaction (QC-PCR). Two recombinant DNA molecules, differing in size due to a 92-bp deletion within the HCMV gH sequence, were used in co-amplification studies to construct a standard curve from which the copy number of the gH gene present in clinical samples could be interpolated. The use of primers labeled with a fluorescent dye allowed direct detection of the amplified products by measuring the amount of fluorescence emitted by each specific PCR fragment with an automated DNA sequencer coupled to a software program. This system was validated subsequently using bronchoalveolar lavage cells obtained from immunocompromised patients and found to be highly sensitive and reproducible over a range of 5-50,000 HCMV gH copies. This rapid procedure could easily be applied to study the pathogenesis of HCMV infection, identify the patients at high risk of developing HCMV disease, and monitor the effects of antiviral therapy at the molecular level.

Isolation of replication-competent molecular clones of visna virus.

Visna virus is the prototypic member of a subfamily of retroviruses responsible for slow infections of animals and humans. As a part of our investigation of the functions of viral gene products in virus replication, we have isolated three infectious molecular clones and determined the complete nucleotide sequences of two of the clones. We have also characterized the progeny of the biologically cloned viral stocks and of the infectious clones and document considerable heterogeneity in plaque size and antigenic phenotype of the former that is reduced to near homogeneity in the progeny of the infectious clones. It thus should now be possible to trace the emergence of antigenic variants of visna virus as well as ascribe defined functions to structural and regulatory genes of the virus in determining neurovirulence and the slow tempo of infection.