Sut-6/NIPP1 modulates tau toxicity.

Neurodegenerative diseases exhibiting the pathological accumulation of tau such as Alzheimer's disease and related disorders still have no disease-modifying treatments and the molecular mechanisms of neurodegeneration remain unclear. To discover additional suppressor of tauopathy (sut) genes that mediate or modulate the toxicity of pathological tau, we performed a classical genetic screen using a tau transgenic Caenorhabditis elegans model. From this screen, we identified the suppressing mutation W292X in sut-6, the C. elegans homolog of human NIPP1, which truncates the C-terminal RNA-binding domain. Using CRISPR-based genome editing approaches, we generated null and additional C-terminally truncated alleles in sut-6 and found that loss of sut-6 or sut-6(W292X) suppresses tau-induced behavioral locomotor deficits, tau protein accumulation and neuron loss. The sut-6(W292X) mutation showed stronger and semi-dominant suppression of tau toxicity while sut-6 deletion acted recessively. Neuronal overexpression of SUT-6 protein did not significantly alter tau toxicity, but neuronal overexpression of SUT-6 W292X mutant protein reduced tau-mediated deficits. Epistasis studies showed tauopathy suppression by sut-6 occurs independent of other known nuclear speckle-localized suppressors of tau such as sut-2, aly-1/aly-3 and spop-1. In summary, we have shown that sut-6/NIPP1 modulates tau toxicity and found a dominant mutation in the RNA-binding domain of sut-6 which strongly suppresses tau toxicity. This suggests that altering RNA-related functions of SUT-6/NIPP1 instead of complete loss of SUT-6/NIPP1 will provide the strongest suppression of tau.

Orthogonal spatial coding with stimulated parametric down-conversion.

Orthogonal optical coding is widely used in classical multi-user communication networks. Using the phase conjugation property of stimulated parametric down-conversion, we extend the current time-domain orthogonal optical coding scheme to the spatial domain to encode and decode image information. In this process, the idler beam inherits the complex conjugate of the field information encoded in the seed beam. An encoding phase mask introduced onto the input seed beam blurs the image transferred to the idler. The original image is restored by passing the coded transferred image through a corrective phase mask placed in the momentum space of the idler beam. We expect that this scheme can also inspire new techniques in secure image transmission, aberration cancellation, and frequency conversion imaging.

Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

Suppression of Nonlinear Optical Rogue Wave Formation Using Polarization-Structured Beams.

A nonlinear self-focusing material can amplify random small-amplitude phase modulations present in an optical beam, leading to the formation of amplitude singularities commonly referred to as optical caustics. By imposing polarization structuring on the beam, we demonstrate the suppression of amplitude singularities caused by nonlinear self-phase modulation. Our results are the first to indicate that polarization-structured beams can suppress nonlinear caustic formation in a saturable self-focusing medium and add to the growing understanding of catastrophic self-focusing effects in beams containing polarization structure.

Generation of volumetrically full Poincaré beams.

Optical communications, remote sensing, particle trapping, and high-resolution imaging are a few research areas that benefit from new techniques to generate structured light. We present a method of generating polarization-structured laser beams that contain both full and partial polarization states. We demonstrate this method by generating an optical beam that contains every state of partial and full polarization. We refer to this beam as a volumetrically full Poincaré beam to distinguish it from full Poincaré beams, which contain all states of full polarization only. In contrast to methods relying upon spatial coherence to generate polarization-structured beams with partial polarization, our method creates well-collimated beams by relying upon temporal coherence.

Measures of body fatness and height in early and mid-to-late adulthood and prostate cancer: risk and mortality in The Pooling Project of Prospective Studies of Diet and Cancer.

BACKGROUND: Advanced prostate cancer etiology is poorly understood. Few studies have examined associations of anthropometric factors (e.g. early adulthood obesity) with advanced prostate cancer risk. PATIENTS AND METHODS: We carried out pooled analyses to examine associations between body fatness, height, and prostate cancer risk. Among 830 772 men, 51 734 incident prostate cancer cases were identified, including 4762 advanced (T4/N1/M1 or prostate cancer deaths) cases, 2915 advanced restricted (same as advanced, but excluding localized cancers that resulted in death) cases, 9489 high-grade cases, and 3027 prostate cancer deaths. Cox proportional hazards models were used to calculate study-specific hazard ratios (HR) and 95% confidence intervals (CI); results were pooled using random effects models. RESULTS: No statistically significant associations were observed for body mass index (BMI) in early adulthood for advanced, advanced restricted, and high-grade prostate cancer, and prostate cancer mortality. Positive associations were shown for BMI at baseline with advanced prostate cancer (HR = 1.30, 95% CI = 0.95-1.78) and prostate cancer mortality (HR = 1.52, 95% CI = 1.12-2.07) comparing BMI ≥35.0 kg/m2 with 21-22.9 kg/m2. When considering early adulthood and baseline BMI together, a 27% higher prostate cancer mortality risk (95% CI = 9% to 49%) was observed for men with BMI <25.0 kg/m2 in early adulthood and BMI ≥30.0 kg/m2 at baseline compared with BMI <25.0 kg/m2 in early adulthood and BMI <30.0 kg/m2 at baseline. Baseline waist circumference, comparing ≥110 cm with <90 cm, and waist-to-hip ratio, comparing ≥1.00 with <0.90, were associated with significant 14%-16% increases in high-grade prostate cancer risk and suggestive or significant 20%-39% increases in prostate cancer mortality risk. Height was associated with suggestive or significant 33%-56% risks of advanced or advanced restricted prostate cancer and prostate cancer mortality, comparing ≥1.90 m with <1.65 m. CONCLUSION: Our findings suggest that height and total and central adiposity in mid-to-later adulthood, but not early adulthood adiposity, are associated with risk of advanced forms of prostate cancer. Thus, maintenance of healthy weight may help prevent advanced prostate cancer.

Quantification of the regional bioarchitecture in the human aorta.

Regional variance in human aortic bioarchitecture responsible for the elasticity of the vessel is poorly understood. The current study quantifies the elements responsible for aortic compliance, namely, elastin, collagen and smooth muscle cells, using histological and stereological techniques on human tissue with a focus on regional heterogeneity. Using donated cadaveric tissue, a series of samples were excised between the proximal ascending aorta and the distal abdominal aorta, for five cadavers, each of which underwent various staining procedures to enhance specific constituents of the wall. Using polarised light microscopy techniques, the orientation of collagen fibres was studied for each location and each tunical layer of the aorta. Significant transmural and longitudinal heterogeneity in collagen fibre orientations were uncovered throughout the vessel. It is shown that a von Mises mixture model is required accurately to fit the complex collagen fibre distributions that exist along the aorta. Additionally, collagen and smooth muscle cell density was observed to increase with increasing distance from the heart, whereas elastin density decreased. Evidence clearly demonstrates that the aorta is a highly heterogeneous vessel which cannot be simplistically represented by a single compliance value. The quantification and fitting of the regional aortic bioarchitectural data, although not without its limitations, including mean cohort age of 77.6 years, facilitates the development of next-generation finite element models that can potentially simulate the influence of regional aortic composition and microstructure on vessel biomechanics.

Introducing mobile fracture prevention services with DXA in Northern Scotland: a comparative study of three rural communities.

Mobile fracture prevention services, with DXA, significantly improved access to care for those at high risk of fracture living in rural areas. Introduction of mobile services facilitated access to fracture liaison services and development of integrated of care pathways across community- and secondary-based care. INTRODUCTION: The ageing population is growing faster in rural areas, yet most fracture prevention services are located in urban areas. As part of a wider study, evaluating the introduction of mobile fracture prevention services, we focus on whether mobile services improve access to care for those at highest risk of fracture. METHODS: Services outcomes were assessed against the Royal Osteoporosis Society clinical standards for fracture liaison services. This included standardised, age-specific referral rates, FRAX 10-year probability of major osteoporotic and hip fracture of referrals, pre- and post-introduction of the mobile service across two island and one rural mainland sites. This was compared with referrals from a similar rural mainland region with local access to a comprehensive service. RESULTS: Greatest impact occurred in areas with most limited service provision at baseline. Mean age of patients referred increased from 59 to 68 years (CI 6.8-10.1, p < 0.001). Referral rates increased from 2.8 to 5.4 per 1000 population between 2011 and 2018, with a 5-fold rise in those ≥ 75 years (0.4 to 2.0 per 1000). Mean FRAX 10-year risk of major osteoporotic fracture increased from 12.7 to 17.7% (CI 3.2-5.7, p < 0.001). Mean hip fracture risk probability increased from 3.0 to 5.7% (CI 2.0-3.4, p < 0.001). However, referral rates from the mobile sites remained lower than the comparator site. CONCLUSIONS: Mobile fracture prevention services, including DXA, greatly improved uptake amongst high-risk individuals. Mobile services facilitated development of integrated of care pathways, including fracture liaison services, across community- and secondary-based care.

Quantum Nonlocal Aberration Cancellation.

Phase distortions, or aberrations, can negatively influence the performance of an optical imaging system. Through the use of position-momentum entangled photons, we nonlocally correct for aberrations in one photon's optical path by intentionally introducing the complementary aberrations in the optical path of the other photon. In particular, we demonstrate the simultaneous nonlocal cancellation of aberrations that are of both even and odd order in the photons' transverse degrees of freedom. We also demonstrate a potential application of this technique by nonlocally canceling the effect of defocus in a quantum imaging experiment and thereby recover the original spatial resolution.

Chiropractic Integrated Care Pathway for Low Back Pain in Veterans: Results of a Delphi Consensus Process.

OBJECTIVE: The purpose of this study was to develop an integrated care pathway for doctors of chiropractic, primary care providers, and mental health professionals who manage veterans with low back pain, with or without mental health comorbidity, within Department of Veterans Affairs health care facilities. METHODS: The research method used was a consensus process. A multidisciplinary investigative team reviewed clinical guidelines and Veterans Affairs pain and mental health initiatives to develop seed statements and care algorithms to guide chiropractic management and collaborative care of veterans with low back pain. A 5-member advisory committee approved initial recommendations. Veterans Affairs-based panelists (n = 58) evaluated the pathway via e-mail using a modified RAND/UCLA methodology. Consensus was defined as agreement by 80% of panelists. RESULTS: The modified Delphi process was conducted in July to December 2016. Most (93%) seed statements achieved consensus during the first round, with all statements reaching consensus after 2 rounds. The final care pathway addressed the topics of informed consent, clinical evaluation including history and examination, screening for red flags, documentation, diagnostic imaging, patient-reported outcomes, adverse event reporting, chiropractic treatment frequency and duration standards, tailored approaches to chiropractic care in veteran populations, and clinical presentation of common mental health conditions. Care algorithms outlined chiropractic case management and interprofessional collaboration and referrals between doctors of chiropractic and primary care and mental health providers. CONCLUSION: This study offers an integrative care pathway that includes chiropractic care for veterans with low back pain.

On the viability of Mg extraction in MgMoN2: a combined experimental and theoretical approach.

Layered MgMoN2 was prepared by solid state reaction at high temperature between Mo and Mg3N2 in N2 which represents a simple synthetic pathway compared to the previously reported method that used NaN3 as the nitrogen source. The crystal structure of MgMoN2 was studied by synchrotron X-ray and neutron powder diffraction. The feasibility of oxidizing this compound and concomitantly extracting magnesium from the structure was assessed by both chemical and electrochemical approaches, using different protocols. The X-ray diffraction patterns of the oxidized samples do not exhibit any relevant difference with respect to that of the as prepared MgMoN2 and no differences in the cell parameters are deduced from Rietveld refinements. No hints pointing at the presence of any amorphous phase are observed either. These results are rationalized through DFT calculated energy barriers for Mg2+ ion migration in MgMoN2.

Comparison of bone healing, as assessed by computed tomography, following tibial tuberosity advancement in dogs with and without autogenous cancellous bone grafts.

AIMS: To objectively compare measures of bone healing, using computed tomography (CT) in dogs following bilateral tibial tuberosity advancement (TTA), between tibiae treated with and without autogenous cancellous bone grafts. METHODS: Ten dogs with bilateral cranial cruciate ligament disease requiring surgical stabilisation were prospectively recruited to undergo single-session bilateral TTA, with only one, randomly assigned, tibia receiving bone graft in the osteotomy deficit. Bone healing at the osteotomy site was assessed using CT performed 38-70 days post-operatively. CT images were evaluated using both objective measurements of osseous bridging and subjective evaluation by six radiologists. Repeated measures ANOVA was used to compare the objective outcomes between the grafted and non-grafted tibiae. RESULTS: The mean percentage of the osteotomy deficit bridged at the lateral cortex was greater in grafted (77.6, SD 35.2%) compared to non-grafted (63.0, SD 36.5%) tibiae (p=0.001), but did not differ at the medial cortex (p=0.1). The mean minimum callus width was greater in grafted (7.2, SD 3.3 mm) compared to non-grafted (3.6, SD 2.9 mm) tibiae (p<0.001). There was no difference in mean attenuation (measured in Hounsfield units) of the callus between grafted and non-grafted tibiae (p=0.5). The grafted tibia was deemed to have superior bone healing in 50/60 subjective assessments made by radiologists. CONCLUSIONS: Superior osseous bridging was detected by CT analysis following TTA using autogenous cancellous bone grafts compared with no graft. This was shown by greater bridging percentage at the lateral cortex and formation of a broader callus. Qualitative assessments made by six radiologists also supported the conclusion that bone healing was improved by use of autogenous cancellous bone graft. CT was a useful method for assessing evidence of bone healing following TTA. CLINICAL RELEVANCE: These findings justify the application of autogenous cancellous bone graft to augment healing following TTA in dogs.

Stochastic interdigitation as a toughening mechanism at the interface between tendon and bone.

Reattachment and healing of tendon to bone poses a persistent clinical challenge and often results in poor outcomes, in part because the mechanisms that imbue the uninjured tendon-to-bone attachment with toughness are not known. One feature of typical tendon-to-bone surgical repairs is direct attachment of tendon to smooth bone. The native tendon-to-bone attachment, however, presents a rough mineralized interface that might serve an important role in stress transfer between tendon and bone. In this study, we examined the effects of interfacial roughness and interdigital stochasticity on the strength and toughness of a bimaterial interface. Closed form linear approximations of the amplification of stresses at the rough interface were derived and applied in a two-dimensional unit-cell model. Results demonstrated that roughness may serve to increase the toughness of the tendon-to-bone insertion site at the expense of its strength. Results further suggested that the natural tendon-to-bone attachment presents roughness for which the gain in toughness outweighs the loss in strength. More generally, our results suggest a pathway for stochasticity to improve surgical reattachment strategies and structural engineering attachments.

Printed three-dimensional airway model assists planning of single-lung ventilation in a small child.

BACKGROUND: Single-lung ventilation in infants and small children is challenging because suitable sizes of double-lumen cuffed tracheal tubes are not available. A 6-yr-old child required pulmonary saline washout for primary alveolar proteinosis, and therefore needed sequential single-lung ventilation in order to achieve safe oxygenation. Before undertaking this potentially hazardous procedure, we practised bronchial intubation on an anatomical model of her airway constructed from computed tomography (CT) data. METHODS: We created a full-scale, anatomically accurate, transparent plastic model of the trachea and main bronchi on a three-dimensional printer using data from a CT scan. We then performed several different airway approaches to identify those likely to be most suitable, ex vivo, before the clinical procedure was carried out on the patient. RESULTS: The model helped us to choose the type and size of bronchial tubes and to practise their insertion beforehand. Subsequently, during anaesthesia, the chosen technique was successful. CONCLUSIONS: Three-dimensional printing of a model of the airway of a small child aided planning of bronchial intubation and single-lung ventilation. Three-dimensional printing of airway structures may have wider application in anaesthesia practice.

Validation of a short questionnaire for estimating dietary calcium intakes.

UNLABELLED: Concern about calcium supplements, and mainly minor side effects (e.g. constipation) impacting on compliance, means that assessing dietary calcium intake is important. There is no suitable biomarker. Compared to food diaries, a short questionnaire was an efficient way of confirming that patients had adequate calcium intakes (>700 or >1,000 mg) INTRODUCTION: Calcium is usually given alongside treatments for osteoporosis, but recent concerns about potential side effects have led to questioning whether supplements are always necessary. It is difficult to assess calcium intake in a clinical setting and be certain that the patient is getting enough calcium. The aim of this study was to determine whether a short questionnaire for estimating dietary calcium intakes in a clinical setting was fit for purpose. METHODS: We assessed dietary calcium intakes using a short questionnaire (CaQ) in patients attending an osteoporosis clinic (n = 117) and compared them with calcium intakes obtained from a 7-day food diary (n = 72) and a food frequency questionnaire (FFQ) (n = 33). RESULTS: Mean (SD) daily calcium intakes from the CaQ were 836 (348) mg; from the diaries, 949 (384) mg; and from the FFQ, 1,141 (387) mg. The positive predictive value (PPV) was >80% for calcium cut-offs > 700 mg and 70% for cut-offs > 1,000 mg. The calcium intakes for the false positives results were not far below the cut-off. For 1,200 mg, the PPV was 67% or less. CONCLUSION: The CaQ is an adequate tool for assessing whether a patient has daily calcium intakes above 700 or 1,000 mg; if below these cut-offs, it is possible that the patient still has enough calcium in the diet, which could be clarified by questioning the patient further. As there were few patients with calcium intakes above 1,200 mg a day, the CaQ cannot be recommended as a tool for confirming higher dietary calcium intakes.

A parallel group double-blind RCT of vitamin D3 assessing physical function: is the biochemical response to treatment affected by overweight and obesity?

UNLABELLED: Vitamin D may affect skeletal muscle function. In a double-blind, randomised, placebo-controlled trial, we found that vitamin D3 supplementation (400 or 1,000 I.U. vs. placebo daily for 1 year with bimonthly study visits) does not improve grip strength or reduce falls. INTRODUCTION: This study aimed to test the supplementation effects of vitamin D3 on physical function and examine associations between overweight/obesity and the biochemical response to treatment. METHODS: In a parallel group double-blind RCT, healthy postmenopausal women from North East Scotland (latitude-57° N) aged 60-70 years (body mass index (BMI), 18-45 kg/m(2)) were assigned (computer randomisation) to daily vitamin D3 (400 I.U. (n = 102)/1,000 I.U. (n = 101)) or matching placebo (n = 102) (97, 96 and 100 participants analysed for outcomes, respectively) from identical coded containers for 1 year. Grip strength (primary outcome), falls, diet, physical activity and ultraviolet B radiation exposure were measured bimonthly, as were serum 25(OH)D, adjusted calcium (ACa) and phosphate. Fat/lean mass (dual energy X-ray absorptiometry), anthropometry, 1,25-dihydroxyvitamin D and parathyroid hormone were measured at baseline and 12 months. Participants and researchers were blinded throughout intervention and analysis. RESULTS: Treatment had no effect on grip strength (mean change (SD)/year = -0.5 (2.5), -0.9 (2.7) and -0.4 (3.3) kg force for 400/1,000 I.U. vitamin D3 and placebo groups, respectively (P = .10, ANOVA)) or falls (P = .65, chi-squared test). Biochemical responses were similar across BMI categories (<25.25-29.99, ≥30 kg/m(2)) with the exception of a small change at 12-months in serum ACa in overweight compared to non-overweight participants (P = .01, ANOVA; 1,000 I.U. group). In the placebo group, 25(OH)D peak concentration change (winter to summer) was negatively associated with weight (r = -.268), BMI (r = -.198), total (r = -.278) and trunk fat mass (r = -.251), with total and trunk fat mass predictive of winter to summer 25(OH)D change (P = .01/.004 respectively, linear regression). CONCLUSION: We found no evidence of an improvement in physical function following vitamin D3 supplementation for 1 year.

Duration of shedding of Verocytotoxin-producing Escherichia coli in children and risk of transmission in childcare facilities in England.

Exclusion of children with presumptive Verocytotoxin-producing Escherichia coli (VTEC) from childcare facilities until negative stool specimens are obtained is routine practice that disrupts families. We estimated the shedding and exclusion duration and transmission risk in such facilities. The study population comprised 225 children aged <6 years attending 201 childcare facilities in England with microbiologically confirmed VTEC in 2010-2011. We estimated the interval from onset to first negative specimen, and identified transmission events with secondary cases linked to facilities. The median duration of shedding was 31 days, and median period of exclusion was 39·5 days. Cases attending facilities while shedding VTEC did so for a median of 2 days before exclusion. Secondary cases occurred in 6/83 facilities (7%) attended by infectious cases. Despite evidence of VTEC shedding at facilities, transmission is relatively low. Revised control guidelines could consider supervised return for prolonged asymptomatic shedders.

Trends in door-to-balloon time and outcomes following primary percutaneous coronary intervention for ST-elevation myocardial infarction: an Australian perspective.

BACKGROUND: Guidelines for patients with ST-elevation myocardial infarction include a door-to-balloon time (DTBT) of ≤90 min for primary percutaneous coronary intervention. AIM: The aim of this study was to assess temporal trends (2006-2010) in DTBT and determine if a reduction in DTBT was associated with improved clinical outcomes. METHODS: We compared annual median DTBT in 1926 STEMI patients undergoing primary percutaneous coronary intervention from the Melbourne Interventional Group registry. ST-elevation myocardial infarction presenting >12 h and rescue percutaneous coronary intervention was excluded. Major adverse cardiac events were analysed according to DTBT (dichotomised as ≤90 min vs >90 min). A multivariable analysis for predictors of mortality (including DTBT) was performed. RESULTS: Baseline demographics, clinical and procedural characteristics were similar in the STEMI cohort across the 5 years, apart from an increase in out-of-hospital cardiac arrest (3.6% in 2006 vs 9.4% in 2010, P < 0.0001) and cardiogenic shock (7.7-9.6%, P = 0.07). The median DTBT (interquartile range) was reduced from 95 (74-130) min in 2006 to 75 (51-100) min in 2010 (P < 0.01). In this period, the proportion of patients achieving a DTBT of ≤90 min increased from 45% to 67% (P < 0.01). Lower mortality and major adverse cardiac event rates were observed with DTBT ≤90 min (all P < 0.01). Multivariable analysis showed that a DTBT of ≤90 min was associated with improved clinical outcomes at 12 months (odds ratio 0.48; 95% confidence interval 0.33-0.73, P < 0.01). CONCLUSION: There has been a decline in median DTBT in the Melbourne Interventional Group registry over 5 years. DTBT of ≤90 min is associated with improved clinical outcomes at 12 months.

First Report of Laurel Wilt Caused by Raffaelea lauricola on Bay Laurel (Laurus nobilis) in the United States.

Bay laurel (Laurus nobilis L.) is an economically important evergreen tree of the family Lauraceae. It is native to Asia Minor and the Balkans and was introduced into the United States for its ornamental and culinary uses (4). In September 2013, a 6-m-tall bay laurel in Gainesville, FL, attracted our attention because it had wilted leaves, discolored sapwood, and ambrosia beetle entrance holes, all symptoms of laurel wilt. In addition, the tree was growing close to an avocado that succumbed to the disease months earlier. In an effort to determine whether the laurel wilt pathogen (Raffaelea lauricola T.C. Harr., Fraedrich & Agaveya) was, indeed, involved in the decline of the tree of current interest, discolored sapwood was sectioned into 5-mm2 pieces, surface disinfested for 30 s in a 4% sodium hypochlorite solution, and plated onto CSMA media (1,2). Within 7 to 14 days, cream-colored, adpressed fungal growth typical of R. lauricola grew from the sapwood pieces (2). DNA was extracted from an isolate of a single conidium (PL1634) and a portion of the 18S rRNA gene was PCR-amplified with primers NS1/NS4, resulting in a 1,021-bp amplicon (GenBank Accession No. KF913344.1), with a BLASTn search revealing 100% homology to several R. lauricola isolates (3). To confirm pathogenicity, six bay laurel seedlings (0.5 m) and a silk bay (0.65 m) (Persea humilis, susceptible control) were wounded twice with a 0.5-mm-diameter drill bit. Then, 30 µl of a spore suspension of PL1634 (1.38 × 105 condia/plant) were introduced into the xylem by pipette and the wounds were wrapped in Parafilm (1). Negative controls consisted of a mock-inoculated (water) and non-inoculated bay laurel plus a mock-inoculated silk bay. Plants were placed in a growth chamber set to a 16/8 h (25/22°C) diurnal light/temperature cycle. After 60 days, all fungal-inoculated plants were completely wilted with dead leaves and subsequent necrosis of stems, while mock- and non-inoculated controls remained asymptomatic. Sapwood dissection revealed xylem discoloration similar to the original infected tree, and fungi morphologically similar to PL1634 were recovered from all inoculated plants upon isolation on CSMA media. Mock- and non-inoculated controls lacked vascular discoloration and fungal growth on media. In order to determine if the redbay ambrosia beetle, Xyleborus glabratus Eichoff (laurel wilt vector) could successfully reproduce in this host, symptomatic branches (7 cm in diameter) of L. nobilis with external evidence of ambrosia beetle attack (frass "toothpicks") were placed in a plastic rearing box within a growth chamber (25°C). Within 4 weeks of incubation, dozens of immature and mature X. glabratus beetles emerged. This is the first record of Koch's postulates being completed for R. lauricola on L. nobilis and the ability of X. glabratus to infest and breed in its stems. This information may be of importance in the event of an introduction of X. glabratus and its fungal associate to Mediterranean areas where bay laurel is either growing wild or being cultivated as valuable commercial crop. References: (1) S. W. Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington et al. Mycotaxon 104:399, 2008. (3) M. A. Innis et al., eds. PCR Protocols: A Guide to Methods and Applications. Academic Press. San Diego, CA, 1990. (4) A. O. Sari et al. New Forest 31:403, 2006.