Insights for disease modeling from single-cell transcriptomics of iPSC-derived Ngn2-induced neurons and astrocytes across differentiation time and co-culture.

BACKGROUND: Trans-differentiation of human-induced pluripotent stem cells into neurons via Ngn2-induction (hiPSC-N) has become an efficient system to quickly generate neurons a likely significant advance for disease modeling and in vitro assay development. Recent single-cell interrogation of Ngn2-induced neurons, however, has revealed some similarities to unexpected neuronal lineages. Similarly, a straightforward method to generate hiPSC-derived astrocytes (hiPSC-A) for the study of neuropsychiatric disorders has also been described. RESULTS: Here, we examine the homogeneity and similarity of hiPSC-N and hiPSC-A to their in vivo counterparts, the impact of different lengths of time post Ngn2 induction on hiPSC-N (15 or 21 days), and the impact of hiPSC-N/hiPSC-A co-culture. Leveraging the wealth of existing public single-cell RNA-seq (scRNA-seq) data in Ngn2-induced neurons and in vivo data from the developing brain, we provide perspectives on the lineage origins and maturation of hiPSC-N and hiPSC-A. While induction protocols in different labs produce consistent cell type profiles, both hiPSC-N and hiPSC-A show significant heterogeneity and similarity to multiple in vivo cell fates, and both more precisely approximate their in vivo counterparts when co-cultured. Gene expression data from the hiPSC-N show enrichment of genes linked to schizophrenia (SZ) and autism spectrum disorders (ASD) as has been previously shown for neural stem cells and neurons. These overrepresentations of disease genes are strongest in our system at early times (day 15) in Ngn2-induction/maturation of neurons, when we also observe the greatest similarity to early in vivo excitatory neurons. We have assembled this new scRNA-seq data along with the public data explored here as an integrated biologist-friendly web-resource for researchers seeking to understand this system more deeply: https://nemoanalytics.org/p?l=DasEtAlNGN2&g=NES . CONCLUSIONS: While overall we support the use of the investigated cellular models for the study of neuropsychiatric disease, we also identify important limitations. We hope that this work will contribute to understanding and optimizing cellular modeling for complex brain disorders.

Sensor-Based Quantification of Peanut Disease Defoliation Using an Unmanned Aircraft System and Multispectral Imagery.

Early leaf spot (Passalora arachidicola) and late leaf spot (Nothopassalora personata) are two of the most economically important foliar fungal diseases of peanut, often requiring seven to eight fungicide applications to protect against defoliation and yield loss. Rust (Puccinia arachidis) may also cause significant defoliation depending on season and location. Sensor technologies are increasingly being utilized to objectively monitor plant disease epidemics for research and supporting integrated management decisions. This study aimed to develop an algorithm to quantify peanut disease defoliation using multispectral imagery captured by an unmanned aircraft system. The algorithm combined the Green Normalized Difference Vegetation Index and the Modified Soil-Adjusted Vegetation Index and included calibration to site-specific peak canopy growth. Beta regression was used to train a model for percent net defoliation with observed visual estimations of the variety 'GA-06G' (0 to 95%) as the target and imagery as the predictor (train: pseudo-R2 = 0.71, test k-fold cross-validation: R2 = 0.84 and RMSE = 4.0%). The model performed well on new data from two field trials not included in model training that compared 25 (R2 = 0.79, RMSE = 3.7%) and seven (R2 = 0.87, RMSE = 9.4%) fungicide programs. This objective method of assessing mid-to-late season disease severity can be used to assist growers with harvest decisions and researchers with reproducible assessment of field experiments. This model will be integrated into future work with proximal ground sensors for pathogen identification and early season disease detection.[Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Interprofessional communication in the hospital setting : a systematic review of the qualitative literature.

Communication plays a key role in the provision of safe patient care, and miscommunication in healthcare can lead to avoidable patient harm or mortality. Interprofessional communication (IPCom) can be challenging due to differences in training, education and roles between healthcare professions. The aim of this systematic review was to synthesize the qualitative evidence regarding healthcare providers' perceptions of interprofessional communication in the hospital setting. Four databases (PubMed, CINAHL, Web of Science, and Embase) were searched for studies that met the inclusion criteria. Eighteen studies were identified as suitable for inclusion in the review and were examined using thematic synthesis. Thematic synthesis led to the development of five descriptive themes: 1) 'Hierarchy", 2) "Interprofessional Ethos," 3) "Healthcare Environment," 4) "Personal Factors" and 5) "Methods of Communication," and two overarching analytical themes: "Barriers to Communication" and "Facilitators to Communication." Personal factors, such as strong interprofessional relationships, were found to be important facilitators to IPCom, while organizational factors, such as challenging and hierarchical working environments, were found to pose barriers to IPCom.

Comparison of the prognostic accuracy of scoring systems, cardiopulmonary exercise testing, and plasma biomarkers: a single-centre observational pilot study.

BACKGROUND: Current approaches to risk assessment before major surgery have important limitations. The aim of this pilot study was to compare predictive accuracy of preoperative scoring systems, plasma biomarkers, and cardiopulmonary exercise testing (CPET) for complications after major non-cardiac surgery. METHODS: Single-centre, observational study of patients aged ≥40 yr undergoing major elective non-cardiac surgery. Before surgery, risk scores were calculated and blood samples collected for measurement of plasma biomarkers. Patients underwent CPET for measurement of anaerobic threshold (AT) and peak oxygen consumption ( peak). After surgery, patients were followed for 28 days to evaluate complications and major adverse cardiac events (MACE). Data are presented as area under the receiver operating characteristic curve (AUROC) with 95% confidence intervals. RESULTS: A total of 100 patients were recruited between April 2009 and October 2010; 17 of whom did not proceed to surgery. CPET variables suggested good predictive accuracy for MACE [AT: AUROC 0.83 (0.69-0.96); peak AUROC 0.81 (0.69-0.96)] and poor predictive accuracy for all complications [AT: AUROC 0.64 (0.52-0.77); peak AUROC 0.64 (0.52-0.77)]. There was a trend towards predictive accuracy of the plasma biomarkers B-type natriuretic peptide and estimated glomerular filtration rate (calculated from serum creatinine) for MACE but not all complications. C-reactive protein, ASA score, and revised cardiac risk index had little or no predictive value. CONCLUSIONS: These pilot data suggest that CPET and plasma biomarkers may improve risk assessment before surgery. Only large clinical studies can confirm this observation and define the optimal use of these tests in clinical practice.

First observation of African tigerfish Hydrocynus vittatus predating on barn swallows Hirundo rustica in flight.

A population of African tigerfish Hydrocynus vittatus from the Schroda Dam, actively prey on barn swallows Hirundo rustica in flight. This behaviour was discovered during a radio telemetry study and documented using a motion picture video camera. These results show that an avivorous diet is a part of the feeding biology of H. vittatus, and may occur in other populations.

Responsivity to criminogenic need in forensic intellectual disability services.

BACKGROUND: Research has shown for some time that addressing criminogenic need is one of the crucial aspects of reducing reoffending in all types of offenders. Criminogenic need such as anger or inappropriate sexual interest is considered to be crucial in the commission of the offence. The aim of the present study is to investigate the extent to which forensic services address the needs of those accepted into services. METHOD: This study reviews the treatment for 197 offenders with intellectual disability accepted into a range of services. Participants' case files were examined to ascertain the extent to which need was addressed through recognised therapies. A standard pro forma was used on which we had established good reliability across four research assistants. RESULTS: The most frequently referred problems were violence and sexual offending. Specialist forensic intellectual disability community services were significantly more likely to provide treatment specifically designed to address index behaviours when compared to generic community services and secure services. CONCLUSIONS: Various possible explanations of these findings are explored including staffing levels, diagnosed mental illness, expertise of staff and clarity of purpose in services.

RENEB Inter-Laboratory Comparison 2021: The Gene Expression Assay.

Early and high-throughput individual dose estimates are essential following large-scale radiation exposure events. In the context of the Running the European Network for Biodosimetry and Physical Dosimetry (RENEB) 2021 exercise, gene expression assays were conducted and their corresponding performance for dose-assessment is presented in this publication. Three blinded, coded whole blood samples from healthy donors were exposed to 0, 1.2 and 3.5 Gy X-ray doses (240 kVp, 1 Gy/min) using the X-ray source Yxlon. These exposures correspond to clinically relevant groups of unexposed, low dose (no severe acute health effects expected) and high dose exposed individuals (requiring early intensive medical health care). Samples were sent to eight teams for dose estimation and identification of clinically relevant groups. For quantitative reverse transcription polymerase chain reaction (qRT-PCR) and microarray analyses, samples were lysed, stored at 20°C and shipped on wet ice. RNA isolations and assays were run in each laboratory according to locally established protocols. The time-to-result for both rough early and more precise later reports has been documented where possible. Accuracy of dose estimates was calculated as the difference between estimated and reference doses for all doses (summed absolute difference, SAD) and by determining the number of correctly reported dose estimates that were defined as ±0.5 Gy for reference doses <2.5 Gy and ±1.0 Gy for reference doses >3 Gy, as recommended for triage dosimetry. We also examined the allocation of dose estimates to clinically/diagnostically relevant exposure groups. Altogether, 105 dose estimates were reported by the eight teams, and the earliest report times on dose categories and estimates were 5 h and 9 h, respectively. The coefficient of variation for 85% of all 436 qRT-PCR measurements did not exceed 10%. One team reported dose estimates that systematically deviated several-fold from reported dose estimates, and these outliers were excluded from further analysis. Teams employing a combination of several genes generated about two-times lower median SADs (0.8 Gy) compared to dose estimates based on single genes only (1.7 Gy). When considering the uncertainty intervals for triage dosimetry, dose estimates of all teams together were correctly reported in 100% of the 0 Gy, 50% of the 1.2 Gy and 50% of the 3.5 Gy exposed samples. The order of dose estimates (from lowest to highest) corresponding to three dose categories (unexposed, low dose and highest exposure) were correctly reported by all teams and all chosen genes or gene combinations. Furthermore, if teams reported no exposure or an exposure >3.5 Gy, it was always correctly allocated to the unexposed and the highly exposed group, while low exposed (1.2 Gy) samples sometimes could not be discriminated from highly (3.5 Gy) exposed samples. All teams used FDXR and 78.1% of correct dose estimates used FDXR as one of the predictors. Still, the accuracy of reported dose estimates based on FDXR differed considerably among teams with one team's SAD (0.5 Gy) being comparable to the dose accuracy employing a combination of genes. Using the workflow of this reference team, we performed additional experiments after the exercise on residual RNA and cDNA sent by six teams to the reference team. All samples were processed similarly with the intention to improve the accuracy of dose estimates when employing the same workflow. Re-evaluated dose estimates improved for half of the samples and worsened for the others. In conclusion, this inter-laboratory comparison exercise enabled (1) identification of technical problems and corrections in preparations for future events, (2) confirmed the early and high-throughput capabilities of gene expression, (3) emphasized different biodosimetry approaches using either only FDXR or a gene combination, (4) indicated some improvements in dose estimation with FDXR when employing a similar methodology, which requires further research for the final conclusion and (5) underlined the applicability of gene expression for identification of unexposed and highly exposed samples, supporting medical management in radiological or nuclear scenarios.

Characterization of peptide aptamers targeting Bfl-1 anti-apoptotic protein.

Bfl-1, an anti-apoptotic protein of the Bcl-2 family, has been identified as a potential therapeutic target for B-cell malignancies. We describe herein the first characterization of peptide aptamers selected against Bfl-1. We show that most of the Bfl-1 peptide aptamers do not interact with Bcl-2, Bcl-xL, or Mcl-1 in yeast and that some of them restore the pro-apoptotic activity of Bax in yeast in which Bax and Bfl-1 proteins are coexpressed. When expressed in mammalian cells, peptide aptamers interact with Bfl-1 and sensitize B-cell lines to apoptosis induced by chemotherapeutic agents. We further demonstrate that a nonconstrained peptide derived from one aptamer variable region reverses Bfl-1 anti-apoptotic activity in HeLa cells through disruption of Bax-Bfl-1 interaction. This peptide also promotes cell death in lymphoma B-cell lines expressing a high level of Bfl-1 and sensitizes these cells to drug-induced apoptosis. Taken together, these results further validate Bfl-1 as a therapeutic target for malignant B-cells and suggest that peptide aptamers may be a useful tool for guiding the identification of small compounds that target the anti-apoptotic Bfl-1 protein.

Epidemiology of Soybean Rust in Soybean Sentinel Plots in Florida.

Since its discovery in the southeastern United States in 2004, soybean rust (SBR) has been variable from year to year. Caused by Phakopsora pachyrhizi, SBR epidemics in Florida are important to understand, as they may serve as an inoculum source for other areas of the country. This study examined the first disease detection date, incidence, and severity of SBR in relation to environmental data, growth stage, and maturity group (MG3, MG5, MG7) in soybean sentinel plots (225 m2) across north Florida from 2005 through 2008. The majority (91%) of the initial infections were observed in MG5 and MG7 soybeans, with plots not becoming infected until growth stage R4 or later. Precipitation was the principle factor affecting disease progress, where disease increased rapidly after rain events and was suppressed during dry periods. On average, plots became infected 30 days earlier in 2008 than 2005. In 2008, there was a significant increase in disease incidence and severity associated with the occurrence of Tropical Storm Fay, which deposited up to 380 mm of rainfall in north Florida. The results of this study indicate that climatic and environmental factors are important in determining the development of SBR in north Florida.

Inter-laboratory comparison of gene expression biodosimetry for protracted radiation exposures as part of the RENEB and EURADOS WG10 2019 exercise.

Large-scale radiation emergency scenarios involving protracted low dose rate radiation exposure (e.g. a hidden radioactive source in a train) necessitate the development of high throughput methods for providing rapid individual dose estimates. During the RENEB (Running the European Network of Biodosimetry) 2019 exercise, four EDTA-blood samples were exposed to an Iridium-192 source (1.36 TBq, Tech-Ops 880 Sentinal) at varying distances and geometries. This resulted in protracted doses ranging between 0.2 and 2.4 Gy using dose rates of 1.5-40 mGy/min and exposure times of 1 or 2.5 h. Blood samples were exposed in thermo bottles that maintained temperatures between 39 and 27.7 °C. After exposure, EDTA-blood samples were transferred into PAXGene tubes to preserve RNA. RNA was isolated in one laboratory and aliquots of four blinded RNA were sent to another five teams for dose estimation based on gene expression changes. Using an X-ray machine, samples for two calibration curves (first: constant dose rate of 8.3 mGy/min and 0.5-8 h varying exposure times; second: varying dose rates of 0.5-8.3 mGy/min and 4 h exposure time) were generated for distribution. Assays were run in each laboratory according to locally established protocols using either a microarray platform (one team) or quantitative real-time PCR (qRT-PCR, five teams). The qRT-PCR measurements were highly reproducible with coefficient of variation below 15% in ≥ 75% of measurements resulting in reported dose estimates ranging between 0 and 0.5 Gy in all samples and in all laboratories. Up to twofold reductions in RNA copy numbers per degree Celsius relative to 37 °C were observed. However, when irradiating independent samples equivalent to the blinded samples but increasing the combined exposure and incubation time to 4 h at 37 °C, expected gene expression changes corresponding to the absorbed doses were observed. Clearly, time and an optimal temperature of 37 °C must be allowed for the biological response to manifest as gene expression changes prior to running the gene expression assay. In conclusion, dose reconstructions based on gene expression measurements are highly reproducible across different techniques, protocols and laboratories. Even a radiation dose of 0.25 Gy protracted over 4 h (1 mGy/min) can be identified. These results demonstrate the importance of the incubation conditions and time span between radiation exposure and measurements of gene expression changes when using this method in a field exercise or real emergency situation.

Brain tumors and epilepsy: pathophysiology of peritumoral changes.

Epilepsy commonly develops among patients with brain tumors, frequently even as the presenting symptom, and such patients consequently experience substantial morbidity from both the seizures and the underlying disease. At clinical presentation, these seizures are most commonly focal with secondary generalization and conventional medical management is often met with less efficacy. The molecular pathophysiology of these seizures is being elucidated with findings that both the tumoral and peritumoral microenvironments may exhibit epileptogenic phenotypes owing to disordered neuronal connectivity and regulation, impaired glial cell function, and the presence of altered vascular supply and permeability. Neoplastic tissue can itself be the initiation site of seizure activity, particularly for tumors arising from neuronal cell lines, such as gangliogliomas or dysembryoblastic neuroepithelial tumors. Conversely, a growing intracranial lesion can both structurally and functionally alter the surrounding brain tissue with edema, vascular insufficiency, inflammation, and release of metabolically active molecules, hence also promoting seizure activity. The involved mechanisms are certain to be multifactorial and depend on specific tumor histology, integrity of the blood brain barrier, and characteristics of the peritumoral environment. Understanding these changes that underlie tumor-related epilepsy may have roles in both optimal medical management for the seizure symptom and optimal surgical objective and management of the underlying disease.

Effect of type of diet and energy intake on milk production of Holstein-Friesian cows with extended lactations.

The aim of this study was to measure the effect of type of diet and level of energy intake on the performance of cows undergoing extended lactations. Ninety-six Holstein-Friesian cows that calved in July and August 2004 were assigned randomly to 1 of 8 groups each of 12 cows (including 4 primiparous cows). Two of the 8 groups were assigned to each of 4 treatments that varied in lactation length (300 or 670 d) and diet (3 diets: control, high, or full total mixed ration (TMR). The 4 treatments were 1) control 300: cows were managed for a 300-d lactation and grazed pasture supplemented with grain and forage to provide a minimum daily dietary intake of 160 MJ of ME/cow; 2) control 670: as for control 300 except that cows were managed for a 670-d lactation; 3) high 670: cows were managed for a 670-d lactation and pasture was supplemented with grain and forage to provide a minimum daily dietary intake of 180 MJ of ME/cow; 4) full TMR 670: cows were managed for a TMR system that included a high body condition score at calving with cows offered a TMR during a 670-d lactation. The TMR was initially offered ad libitum indoors until about 440 DIM when the amount of TMR offered was reduced by about 2 kg of DM/d to prevent excessive weight gain. The proportions of cows still milking at the end of a 670-d lactation were similar for the control and high dietary groups. The full TMR group had fewer cows milking at 600 DIM: 17 cows milking compared with 24 cows in the control 670 group and 22 cows in the high 670 group. For the period 1 to 670 DIM, increasing the energy level in the diet (control 670 vs. high 670) resulted in a similar yield of milk and a similar fat concentration in the milk, but greater yields of milk fat and protein and greater milk protein percentage of the milk. The full TMR 670 group produced greater yields of milk and milk components (fat, protein, and lactose) and also protein percentage in the milk than the other groups. The milk solids (fat + protein) ratio for the 3 extended-lactation groups, defined as production achieved during the 24-mo calving interval divided by 2 yr (annualized production) expressed as a ratio of that produced in the normal 12-mo calving interval, was not affected by increasing the level of grain in the pasture-based diets (0.93 vs. 0.90 for control and high diets, respectively), but decreased with the TMR diet (0.79). The control 670 group produced 7.1% less milk, but only 2.4% less milk solids than the control 300 group over the 2-yr period of the study. Combining our data with that from 2 earlier studies of extended lactation demonstrated that Holstein cows with a high proportion of Northern Hemisphere genes offered pasture-based diets could achieve a high milk solids ratio, a greater proportion of cows milking at drying-off, and lower body weight gain over the lactation.

Where next for youth mental health services in Ireland?

To date, Ireland has been a leading light in the provision of youth mental health services. However, cognisant of the efforts of governmental and non-governmental agencies working in youth mental health, there is much to be done. Barriers into care as well as discontinuity of care across the spectrum of services remain key challenges. This editorial provides guidance for the next stage of development in youth mental care and support that will require significant national engagement and resource investment.

Downregulation of Bfl-1 protein expression sensitizes malignant B cells to apoptosis.

Elevated expression of the antiapoptotic protein Bfl-1 (A1) was previously reported in several cancer cell lines. Recently, molecular profiling of large B-cell lymphoma identified Bfl-1 as a gene signature in 'OxPhos' diffuse large B-cell lymphoma subtype and in primary mediastinal large B-cell lymphoma, suggesting that in addition to Bcl-2, Bcl-xL and Mcl-1, Bfl-1 may be a relevant target in the design of new strategies for cancer therapy. Using short hairpin RNA strategy, we show here that Bfl-1 silencing in one lymphoblastoid B-cell line and in two diffuse large B-cell lymphoma cell lines potently induces their apoptosis and sensitizes those cell lines to anti-CD20 (Rituximab)-mediated cell death as well as to apoptosis induced by chemotherapeutic molecules such as doxorubicin, vincristine, cisplatin and fludarabine. These results demonstrate for the first time that Bfl-1 is an essential protein for survival of malignant B cells and suggest Bfl-1 may represent a potential target for future drug development against B cell lymphoma.

Modifiable risk factors for intracerebral hemorrhage: study of anticoagulated patients.

OBJECTIVE: To determine whether there are modifiable risk factors for spontaneous intracerebral hemorrhage in patients receiving oral anticoagulation (OAC) therapy. DESIGN: Retrospective chart review between January 2002 and December 2004. PARTICIPANTS: A total of 315 consecutive patients presenting with spontaneous intracerebral hemorrhage. MAIN OUTCOME MEASURES: Overall mortality rates and surgical mortality rates, and discharge home compared with discharge to a long-term care facility. RESULTS: Of the 315 patients reviewed, 65 (20.6%) were receiving OAC therapy. Age, Glasgow Coma Scale score, and size of hematoma at presentation were similar in the 65 patients taking OAC and the 250 patients not taking it. Mean arterial pressure at presentation was significantly higher in the OAC group than in the control group (132 mm Hg vs 107 mm Hg, P = .01) as was the number of hematomas that progressed (52% vs 14%, P = .01). Overall mortality rates were higher in the OAC group than in the control group (52% vs 41%, P = .03) as were surgical mortality rates (62% vs 41%, P = .04). There were no significant differences in morbidity between the 2 groups. CONCLUSION: Mortality rates were higher among patients taking OAC therapy despite their having similarly sized hematomas at presentation. The higher initial mean arterial pressure among such patients has not been described previously in this setting. This higher mean arterial pressure correlates with the propensity of these patients' hematomas to expand after initial imaging and might partially mediate the mortality effect. In patients taking OAC, hypertension appears to be a modifiable risk factor for morbidity and mortality from intracerebral hemorrhage.

Routine use of PICO dressings may reduce overall groin wound complication rates following peripheral vascular surgery.

BACKGROUND: Approximately 19% of morbidity in peripheral vascular surgery is attributable to wound complications, which can result in delayed healing, and also arterial or graft infection leading to limb loss and even mortality in extreme cases. AIM: To determine whether groin wound complications were reduced following the routine introduction of PICO negative pressure wound therapy dressings in patients who underwent peripheral vascular surgery. METHODS: Patients who underwent peripheral vascular surgery from 2011 to 2016 were identified and divided into PICO and non-PICO groups. Patient, procedure and wound characteristics were tabulated and analysed. Patients were followed-up for at least six weeks postoperatively. Wound complication rates, infection confirmed by microbiology, and requirement for re-admission due to wound complications were noted. Basic cost analysis was performed. FINDINGS: In total, 151 patients were analysed (N = 73 PICO, N = 78 non-PICO). No difference in age (P = 0.862), body mass index (P = 0.673), diabetes (P = 0.339), pre-operative albumin (P = 0.196), use of drain (P = 0.343) and history of meticillin-resistant Staphylococcus aureus (P = 0.281) was observed between groups. The PICO group contained more smokers than the non-PICO group (45% vs 29%, P = 0.034). Wound complications were seen in 8% (N = 6) of the PICO group and 19% (N = 15) of the non-PICO group (P = 0.042). No significant difference in infection was found between the two groups (3% vs 6%, P = 0.249), but fewer seromas were observed when PICO dressings were used (1.4% vs 7.7%, P = 0.069). Haematoma (2.7% vs 3.8%, P = 0.531) and dehiscence rates (1.4% vs 1.3%, P = 0.735) were similar between the two groups. CONCLUSIONS: Routine use of PICO dressings is associated with a reduction in wound complication rates following peripheral vascular surgery, and is cost-effective.

Validating Baboon Ex Vivo and In Vivo Radiation-Related Gene Expression with Corresponding Human Data.

The research for high-throughput diagnostic tests for victims of radio/nuclear incidents remains ongoing. In this context, we have previously identified candidate genes that predict risk of late-occurring hematologic acute radiation syndrome (HARS) in a baboon model. The goal of the current study was to validate these genes after radiation exposure in humans. We also examined ex vivo relative to in vivo measurements in both species and describe dose-response relationships. Eighteen baboons were irradiated in vivo to simulate different patterns of partial- or total-body irradiation (TBI), corresponding to an equivalent dose of 2.5 or 5 Sv. Human in vivo blood samples were obtained from patients exposed to different dose ranges: diagnostic computerized tomography (CT; 0.004-0.018 Sv); radiotherapy for prostate cancer (0.25-0.3 Sv); and TBI of leukemia patients (2 × 1.5 or 2 × 2 Sv, five patients each). Peripheral whole blood of another five baboons and human samples from five healthy donors were cultivated ex vivo and irradiated with 0-4 Sv. RNA was isolated pairwise before and 24 h after irradiation and converted into cDNA. Gene expression of six promising candidate genes found previously by us in a baboon model ( WNT3, POU2AF1, CCR7, ARG2, CD177, WLS), as well as three genes commonly used in ex vivo whole blood experiments ( FDXR, PCNA, DDB2) was measured using qRT-PCR. We confirmed the six baboon candidate genes in leukemia patients. However, expression for the candidate gene FDXR showed an inverse relationship, as it was downregulated in baboons and upregulated in human samples. Comparisons among the in vivo and ex vivo experiments revealed the same pattern in both species and indicated peripheral blood cells to represent the radiation-responsive targets causing WNT3 and POU2AF1 gene expression changes. CCR7, ARG2, CD177 and WLS appeared to be altered due to radiation-responsive targets other than the whole blood cells. Linear dose-response relationships of FDXR, WNT3 and POU2AF1 using human ex vivo samples corresponded with human in vivo samples, suggesting that ex vivo models for in vivo dose estimates can be used over a wide dose range (0.001-5 Sv for POU2AF1). In summary, we validated six baboon candidate genes in humans, but the FDXR measurements underscored the importance of independent assessments even when candidates from animal models have striking gene sequence homology to humans. Since whole blood cells represented the same radiation-responsive targets for FDXR, WNT3 and POU2AF1 gene expression changes, ex vivo cell culture models can be utilized for in vivo dose estimates over a dose range covering up to 3.5 log scales. These findings might be a step forward in the development of a gene expression-based high-throughput diagnostic test for populations involved in large-scale radio/nuclear incidents.

Clinical predictors of facial nerve outcome after translabyrinthine resection of acoustic neuromas.

PURPOSE: The translabyrinthine approach to acoustic neuroma resection offers excellent exposure for facial nerve dissection with 95% preservation of anatomic continuity. Acceptable outcome in facial asymptomatic patients is reported at 64-90%, but transient postoperative deterioration often occurs. The objective of this study was to identify preoperative clinical presentation and intraoperative surgical findings that predispose patients to facial nerve dysfunction after acoustic neuroma surgery. METHODS: The charts of 128 consecutive translabyrinthine patients were examined retrospectively to identify new clinical and intraoperative predictors of facial nerve outcome. Postoperative evaluation of patients to normal function or mild asymmetry upon close inspection (House-Brackmann grades of I or II) was defined as an acceptable outcome, with obvious asymmetry to no movement (grades III to VI) defined as unacceptable. Intraoperative nerve stimulation was performed in all cases, and clinical grading was performed by a single neurosurgeon in all cases. RESULTS: Among patients with no preoperative facial nerve deficit, 87% had an acceptable result. Small size (P < 0.01) and low intraoperative nerve stimulation of < 0.10 mA (P< 0.01) were reaffirmed as predictive of functional nerve preservation. Additionally, preoperative tinnitus (P = 0.03), short duration of hearing loss (P< 0. 01), and lack of subjective tumour adherence to the facial nerve (P = 0.02) were independently correlated with positive outcome. CONCLUSIONS: Our experience with the translabyrinthine approach reveals the previously unestablished associations of facial nerve outcome to include presence of tinnitus and duration of hypoacusis. Independent predictors of tumour size and nerve stimulation thresholds were reaffirmed, and the subjective description of tumour adherence to the facial nerve making dissection more difficult appears to be important.

Anti-cancer properties of phenolics from apple waste on colon carcinogenesis in vitro.

Colorectal cancer is one of the most common cancers in Western countries. The World Health Organisation identifies diet as a critical risk factor in the development and progression of this disease and the protective role of high levels of fruit and vegetable consumption. Several studies have shown that apples contain several phenolic compounds that are potent anti-oxidants in humans. However, little is known about other beneficial properties of apple phenolics in cancer. We have used the HT29, HT115 and CaCo-2 cell lines as in vitro models to examine the effect of apple phenolics (0.01-0.1% apple extract) on key stages of colorectal carcinogenesis, namely; DNA damage (Comet assay), colonic barrier function (TER assay), cell cycle progression (DNA content assay) and invasion (Matrigel assay). Our results indicate that a crude extract of apple phenolics can protect against DNA damage, improve barrier function and inhibit invasion (p<0.05). The anti-invasive effects of the extract were enhanced with twenty-four hour pretreatment of cells (p<0.05). We have shown that a crude apple extract from waste, rich in phenolic compounds, beneficially influences key stages of carcinogenesis in colon cells in vitro.

Syndromic and sporadic pediatric optic pathway gliomas: review of clinical and histopathological differences and treatment implications.

Optic pathway gliomas (OPGs) are the most common primary neoplasm of the optic pathway. These lesions usually present in childhood and can arise anywhere along the optic pathway; they occur more frequently in women; and they rarely undergo late progression. Management strategies after the initial diagnosis are controversial, compounded by the different behaviors exhibited by sporadic and syndromic tumors. Neurofibromatosis Type 1 (NF1), with aberrant oncogenic signaling and consequent predisposition to intracranial tumors, is the most common associated syndrome, with nearly 20% of NF1 patients developing OPGs. A comorbid NF1 diagnosis has implications for tumor location with greater predilection for optic nerve involvement, whereas chiasmal and postchiasmal lesions are more frequently seen in sporadic cases. Syndromic OPGs often exhibit more indolent behavior and lower rates of clinical progression, and the majority of these are diagnosed by routine neuroophthalmological screening. When treatment is indicated, however, the molecular abnormalities that constitute this syndrome can limit the available chemotherapy and radiotherapy options because clinicians fear secondary malignancy and cerebrovascular complications. Furthermore, radiotherapy early in life can impair an individual's intellectual development, endocrine function, and physical growth, thereby limiting the role of this modality in the treatment of this childhood lesion. Differential gene expression and histogenesis among sporadic and syndromic OPGs may account for the different tumor behaviors, but studies correlating specific genetic and proteomic changes with patient outcome are pending. Loss of heterozygosity at 10 and 17q are more common among patients with NF1, and Ki67 labeling intensity of 2-3% and low p53 labeling intensity seem prognostic of aggressive tumor behavior. Recent advances in the development of a preclinical mouse model of NF1-associated OPG will permit investigation into improved detection strategies and chemotherapeutic and radiotherapy treatment protocols.