Enhanced production of antifungal lipopeptides by Bacillus amyloliquefaciens for biocontrol of postharvest disease.

Food security to sustain increasing populations is a global concern. A major factor threatening food security is crop spoilage during postharvest storage. Reduction of postharvest spoilage has mainly been addressed by the application of synthetic chemicals. Bacillus lipopeptides, specifically lipopeptide homologues exhibiting antifungal efficacy, offer an alternative environmentally benign protocol for reduction of postharvest phytopathogens. This work is directed towards Bacillus lipopeptide production for biocontrol of postharvest phytopathogens in general and fungal phytopathogens in particular. Bacillus amyloliquefaciens DSM 23117 was identified as an organism with superior potential for lipopeptide production, via screening of 4 Bacillus candidates, in terms of antifungal lipopeptide concentration, yield, productivity and preferred homologue ratio. Efficacy of B. amyloliquefaciens lipopeptides against Botrytis cinerea substantiated appropriateness of this Bacillus species. Subsequent process modification of B. amyloliquefaciens cultures demonstrated that the concentration and ratio of the lipopeptides were significantly influenced by process conditions and further, distinguished nitrate and oxygen availability as key parameters defining optimal lipopeptide production. Discrete B. amyloliquefaciens cultures supplied with 4, 8, 10 and 12 g/L NH4NO3 demonstrated optimal lipopeptide concentration, yield and productivity, with respect to both total and antifungal lipopeptides, in the culture containing 8 g/L NH4NO3. Enhancement of total and antifungal lipopeptide kinetics similar to those quantified on increasing the nitrate from 4 to 8 g/L NH4NO3 were exhibited in B. amyloliquefaciens cultures when the oxygen in the sparge gas was increased from 21 to 30 mol%. The enhancement of lipopeptide production under conditions of increased nitrate and increased oxygen supply is explained in terms of increased availability of nitrogen for synthesis. This work has highlighted key parameters for maximisation of Bacillus lipopeptide production and manipulation of antifungal/surfactin ratios for optimum efficacy and informs on future development of process strategies towards production optimisation of antifungal lipopeptides as a green alternative to synthetic chemicals.

The utilization of circulating cell-free fetal DNA testing and decrease in invasive diagnostic procedures: an institutional experience.

OBJECTIVE: To characterize the patient population utilizing circulating cell-free fetal DNA (ccffDNA) testing at a large academic center and evaluate trends in the performance of invasive diagnostic procedures. STUDY DESIGN: A retrospective cohort study of all patients who underwent cell-free DNA testing from May to December 2012 was performed. RESULT: During the study period, 206 patients had cell-free DNA testing. Of those, 75% (155/206) were of ages ⩾ 35 years. Of those undergoing ccffDNA testing, 41% had positive aneuploidy screening and 38% had abnormal ultrasound findings. Only 7% of the patients with negative ccffDNA testing opted for an invasive diagnostic procedure compared with 60% with positive testing (P<0.01). The rate of invasive procedures decreased from 5.9% of all visits to the center during a similar 8-month period in 2010 to 4.1% of all visits during the study period (P<0.01). CONCLUSION: Our data suggest that ccffDNA testing leads to reduced uptake of invasive procedures.

Barbiturate ingestion in three adult captive tigers (Panthera tigris) and concomitant fatal botulism of one.

Zoo animals, including tigers, have been reported to suffer from barbiturate intoxication, with pentabarbitone being most commonly recorded. Clinical signs range from mild ataxia to general anaesthesia with recovery over hours to days with several factors affecting hepatic barbiturate metabolism and tissue partitioning. Botulism is an often fatal intoxication in man, animals, birds and certain fish. The occurrence in carnivores is uncommon to rare, with only 2 reports found of botulism in felids. This report relates to 3 adult captive cohabiting tigers that simultaneously developed signs of abdominal discomfort, progressive ataxia, recumbency and comatose sleep resembling stage 2 anaesthesia, alternating with periods of distracted wakefulness and ataxic movements. These signs occurred 4 days after being fed the carcass of a horse that had ostensibly died of colic and not been euthanased. The male tiger that was the dominant animal in the feeding hierarchy was worst affected and had to be given intravenous fluids. The female that was lowest in hierarchy was unaffected. After 48-72 hours of treatment at the Onderstepoort Veterinary Academic Hospital the females could eat and made an uneventful recovery. The male tiger showed partial recovery but died during the night a few hours after drinking water on his return to the owner. Necropsy revealed severe oesophageal dilation and impaction with decaying grass; some of this material and water were present in the pharynx and trachea, and had been aspirated causing acute widespread bronchopneumonia. Colon content tested negative for common pesticides but, together with liver, tested positive for barbiturate. Serum taken on the day of admission had tested negative for barbiturate and the residual serum from the 3 animals later tested negative for botulinum toxin. Colon and oesophageal content from the male at necropsy were positive for Clostridium botulinum toxin type C by the mouse bioassay neutralisation test, confirming that this male had had concomitant barbiturate toxicity and botulism, and had succumbed to aspiration bronchopneumonia secondary to pharyngeal, laryngeal and oesophageal paralysis and oesophageal

Personality Profile and Coping Resources of Family Medicine Vocational Trainees at the University of Limpopo; South Africa

Background: Doctors are exposed to various stress factors in their personal and family lives; as well as in the workplace. Stress inherent to the responsibilities and challenges of the medical field may become a health hazard and threaten the well-being of the medical practitioner. Methods: The aim of this study was to investigate the personality traits and coping resources that contribute to the wellbeing of medical practitioners. A cross-sectional study of 44 out of 45 (98response rate) family medicine vocational trainees at the Medical University of Southern Africa (now known as the University of Limpopo) was conducted. A biographic questionnaire was utilised to obtain specific information regarding the participants. The principal researcher used the Coping Resources Inventory (CRI) questionnaire to assess coping resources; and the 16PF personality analysis (16PF) to establish a personality profile of the participants. Results: The majority of participants (81.8) indicated that they mainly experienced work-related stress. Thirty-two participants (72.72) self-medicated. Fourteen participants (31.81) claimed to experience burn-out and twenty (45.45) reported fatigue. In terms of their coping resources; 24 male participants (54.54) did not cope socially (p . 0.008) and eight (18.18) also did not cope physically (p . 0.024). Conclusions: The medical practitioners had a universal personality profile. They lacked insight regarding the symptoms they were experiencing that warranted management; e.g. depression and anxiety. The medical practitioners in this study did not utilise their social and physical coping resources optimally and reported poor help-seeking behaviour

Transperineal three-dimensional ultrasound in evaluating anal sphincter muscles.

OBJECTIVES: To evaluate whether transperineal three-dimensional (3D) ultrasound can be used to depict normal anal sphincter anatomy and to measure the thickness of muscle layers and the anteroposterior length of the levator hiatus. METHODS: The study included 22 normal nulliparous female volunteers. Transperineal 3D sonographic evaluation of the anal canal included assessment of sphincter shape, echogenicity, marginal definition and muscle thickness. Measurements of the thickness of the internal anal sphincter (IAS) and puborectalis muscle (PRM) were determined with the women at rest and during squeezing by two observers, and interobserver reliability was determined. The anteroposterior length of the levator hiatus at rest and during squeezing was measured. RESULTS: The proximal end of the anal canal (towards the rectum) consisted of overlapping IAS and PRM, and the distal end (towards the anus) consisted of overlapping IAS and external anal sphincter (EAS). At the PRM level, the mean +/- SD IAS thickness was 2.3 +/- 0.5 mm at rest and 2.5 +/- 0.4 mm during squeezing, and at the mid-EAS level it was 2.9 +/- 0.5 mm at rest and 2.8 +/- 0.5 mm during squeezing. The PRM thickness was 6.5 +/- 1.0 mm at rest and 6.4 +/- 1.2 mm during squeezing. The difference in muscle thickness of the sphincter layers with the woman at rest and during squeezing was not significant. The anteroposterior length of the levator hiatus was 51.7 +/- 5.0 mm at rest and 47.4 +/- 4.1 mm during squeezing (P < 0.01). CONCLUSION: Transperineal 3D ultrasound may be useful in evaluating the anatomy of the anal canal.

Standardization of three-dimensional images in obstetrics and gynecology: consensus statement.

Standardization of the display of ultrasound images has so far only been achieved in transabdominal two-dimensional (2D) sonography. In contrast, there is a lack of uniformity in the demonstration of transvaginal 2D ultrasound images. The described non-uniformity frequently leads to confusion in the assessment of an image, in particular with regard to the accurate anatomical assignment of left/right and dorsal/ventral. Three-dimensional (3D) sonography offers a unique opportunity to avoid this confusion in the interpretation of ultrasound images, because, independent of primary volume acquisition, the volume can always be rotated so that the stored object can at all times be visualized in a known anatomical position, rendering it of no importance whether the image acquired transvaginally is demonstrated from above or from below. This will also be important in allowing fusion of ultrasound image data with computed tomographic, magnetic resonance and/or positron emission tomography images. In this article we suggest that standardization of transabdominal and transvaginal 3D images does not only provide the inexperienced physician/sonographer with a guide to spatial orientation, but also serves to avoid erroneous topographical interpretations.

Prenatal three-dimensional ultrasound: perception of sonographers, sonologists and undergraduate students.

OBJECTIVE: To assess the perception of non-pregnant sonographers, sonologists and undergraduate students on the use of three-dimensional (3D) ultrasound technology in fetal medicine. METHODS: This was a study of two groups of non-pregnant subjects. Group I included 520 (305 female, 215 male) medical professionals who completed a questionnaire after attending a lecture on 3D imaging. Factors such as gender, career and having children were analyzed with respect to the attendee's responses about use of 3D ultrasound for medical purposes and for reassurance. Group II included 137 (75 female, 60 male, two unknown) undergraduate students from bioengineering, psychology and physiology classes who completed a questionnaire after attending a brief presentation on two-dimensional and 3D fetal imaging. Factors such as gender and area of educational interest were analyzed with respect to the students' responses about the use of 3D ultrasound for medical purposes and for parental-fetal attachment. RESULTS: In Group I, 63% said that they would like to have a 3D ultrasound examination in the future, while 14% said that they would not. Common reasons given for wanting a 3D ultrasound exam in the future were for medical purposes (39%) or reassurance (18%). The main differences perceived between two-dimensional (2D) and 3D ultrasound were medical advantages (65%) and parental reassurance (28%). 62.4% of Group I thought 3D technology should be in wide use in obstetric ultrasound and 73.6% thought that 3D ultrasound would reassure parents carrying normal fetuses. Gender, age and career did not have a significant influence on perception of 3D ultrasound. In Group II, the majority (91%) said they could see a remarkable difference between 2D and 3D ultrasound. 83% responded that they would like to have a 3D ultrasound examination of their own baby in the future for the following reasons: 34% for the detailed picture, 31% for increased abnormality detection, 13% for reassurance or curiosity; 8% thought it would be unnecessary or a negative experience. Concerning parental-fetal attachment, 72% thought 3D ultrasound would have a positive effect. The majority of Group II (93%) thought 3D ultrasound would be valuable and 56% thought 3D ultrasound would assist in diagnosing fetal abnormalities. There was no significant relationship between gender, age or area of interest and the perception of 3D ultrasound. CONCLUSIONS: Responses by sonographers and physicians suggest that 3D ultrasound will have a role in the future for medical indications and in reassuring patients carrying normal fetuses. Our results also suggest that undergraduate students believe that 3D ultrasound will be a valuable technique in obstetrics and that it will have a positive effect on parental-fetal attachment.

Prenatal detection of associated anomalies in fetuses diagnosed with cleft lip with or without cleft palate in utero.

OBJECTIVE: The aim of this study was to determine the prenatal detection rate of associated anomalies in fetuses with a suspected cleft lip with or without cleft palate. METHODS: Fetuses with a suspected cleft lip with or without cleft palate, determined by prenatal ultrasound, were prospectively enrolled. Additional anomalies suspected by ultrasound or genetic testing were recorded. Postnatal outcome was obtained. RESULTS: Forty-five fetuses with a cleft lip with or without cleft palate, diagnosed prenatally with either two-dimensional and/or three-dimensional ultrasound, were studied. Postnatal follow-up revealed that 16 (35.6%) of these 45 fetuses had an additional structural or syndromic abnormality. Of the 37 fetuses with prenatally determined 'isolated' cleft lip with or without cleft palate, eight (21.6%) had an additional malformation identified after delivery. CONCLUSION: In pregnancies complicated by a cleft lip with or without cleft palate, patients should be informed of the risks of associated anomalies, some of which may be undetected prenatally.

Effects of ultrasound on maternal-fetal bonding: a comparison of two- and three-dimensional imaging.

OBJECTIVES: The purpose of this study was to evaluate the effect of two-dimensional (2DUS) compared to three-dimensional ultrasound (3DUS) imaging on the maternal-fetal bonding process. METHODS: Fifty mothers who had 2DUS and 50 who had 2DUS and 3DUS were included in the study. A postpartum survey by telephone interview was carried out to assess maternal-fetal bonding. Bonding was evaluated by analysis of extent of prenatal image sharing, maternal ability to form a mental picture of the baby and mother's comments about their ultrasound images. Data were analyzed using the independent t-test, Chi-square and Mann-Whitney U-tests. RESULTS: Mothers who received 3DUS showed their ultrasound images to more people (median, 27.5; interquartile range, 14.5-40.0) than mothers receiving 2DUS alone (median, 11.0; interquartile range, 5.0-25.5) (P < 0.001, Z = -3.539). Eighty-two percent of the subjects screened with 3DUS had a greater tendency to form a mental picture of the baby postexamination compared to 39% of the 2DUS subjects (P < 0.001, Z = -3.614). Mothers receiving a 3DUS study were more likely to receive comments on the similarities/differences of the neonate compared to those having 2DUS studies. Furthermore, 70% of the mothers receiving 3DUS felt they 'knew' the baby immediately after birth vs. 56% of the mothers receiving 2DUS (P = 0.009, Z = -2.613). Both 2DUS and 3DUS experiences were positive, however, the comments made by the mothers undergoing 3DUS (n = 18) were more exclamatory (amazed, wonderful, fabulous) than those undergoing 2DUS (n = 4). Patients having a 3DUS examination consistently scored higher than those having a 2DUS examination alone for all categories of maternal-fetal bonding. CONCLUSION: 3DUS appears to more positively influence the perceptions of mothers to their babies postbirth compared to 2DUS. Specifically, mothers who had 3DUS showed their ultrasound images to a greater number of people compared to mothers who had 2DUS alone and this may represent mother's social support system. 3DUS may have a greater impact on the maternal-fetal bonding process.

Prenatal screening for congenital heart disease using real-time three-dimensional echocardiography and a novel 'sweep volume' acquisition technique.

OBJECTIVES: Conventional prenatal screening for congenital heart disease (CHD) involves a time-consuming and highly operator-dependent acquisition of the four-chamber view and outflow tracts. By acquiring the entire fetal heart instantaneously as a single volume, real-time three-dimensional echocardiography (RT3DE) may facilitate fetal cardiac screening. METHODS: Four reviewers, each experienced with fetal cardiac imaging, blindly and independently evaluated a single cardiac volume from each of 18 fetuses (11 normal, seven with CHD). Two-dimensional echocardiography served as the gold standard. Three-dimensional evaluation of each fetus included a series of volume acquisitions lasting 2-6 s each. A 'sweep volume' technique was developed to fit larger hearts into a single non-gated volume. RESULTS: RT3DE had a high sensitivity for detecting CHD (93%), with only a single case being missed by two observers. Specificity for CHD was low (45%), with a high rate of 'cannot determine' responses and false positive artifacts. CONCLUSIONS: These preliminary results suggest that RT3DE has the potential to function as a screening tool for fetal heart disease. However, artifacts must be recognized and minimized, resolution must improve, and substantial training will be necessary prior to widespread clinical use.

Potential of three-dimensional ultrasound in neonatal and paediatric neurosonography.

The aim of this study was to describe the potential of three-dimensional ultrasound (3D US) in paediatric and neonatal neurosonography. The potential applications are illustrated based on our experience in 150 patients using three different 3D US techniques at two different sites. Various disease entities throughout the paediatric age have been evaluated. The potential of 3D US, including 3D US of the cerebral vessels based on colour Doppler data, is discussed based on comparison with conventional 2D US or other imaging (as available), and with regard to the literature. In our experience, 3D US is feasible in neonatal and paediatric neurosonography. It reduces imaging time, improves demonstration of complex anatomy and vasculature, and allows for evaluation of anatomy/pathology in any plane. The 3D US furthermore improves volume assessment (e.g. in hydrocephalus), and comparison with CT, MRI and during follow-up, with a potentially improved standardisation and documentation. The 3D US additionally offers an ideal modality for training and education, as the brain and the neonatal spine can be virtually rescanned at the workstation. Yet, limitations such as areas inaccessible to 2D US, limited resolution and motion artefacts have to be acknowledged. Three-dimensional US has the potential to become a valuable additional imaging tool in paediatric neurosonography.

Feasibility of performing a virtual patient examination using three-dimensional ultrasonographic data acquired at remote locations.

OBJECTIVE: To evaluate the feasibility of performing three-dimensional ultrasonographic studies that meet American Institute of Ultrasound in Medicine and American College of Radiology ultrasonographic examination guidelines with review off-line and at remote locations. METHODS: One hundred patients were studied at 2 institutions using high-end two-dimensional clinical ultrasonographic scanners and commercially available three-dimensional ultrasonography for a variety of organ systems (first- and second-trimester fetus, abdomen, and female pelvis). We evaluated several parameters, including measurements, completeness of organ visualization, abnormalities identified, image quality, number of volumes required, and discrepancies between interpretations. RESULTS: Overall, three-dimensional ultrasonography could produce diagnostic-quality results comparable with those of two-dimensional ultrasonography. Three-dimensional ultrasonographic image quality was lower than that of two-dimensional ultrasonography. Two- and three-dimensional ultrasonographic measurements were comparable (<5% difference), as was the extent of organ visualization, although some structures were challenging for both two- and three-dimensional ultrasonography. In general, organs completely imaged in the scanner field of view required 1 to 1.5 volumes, whereas larger organs required between 3 and 6 volumes. Differences among reviewers' interpretations highlighted the need for standardization of acquisition and reviewing protocols for sonographers and physicians. CONCLUSIONS: Our results show that it is clinically feasible to acquire three-dimensional ultrasonographic data at one site and to obtain accurate interpretation by off-line review at another within the context of providing high-quality clinical diagnostic studies.

Three-dimensional ultrasound in obstetrics and gynecology.

Three-dimensional ultrasound is a new modality finding its way into clinical practice. Most of the major ultrasound vendors are now developing three-dimensional ultrasound capabilities. We expect that although three-dimensional ultrasound will not replace two-dimensional ultrasound, many additional benefits will be identified and its use will continue to grow. The ability to evaluate anatomy and pathology with multiplanar and surface-rendered images provides physicians additional valuable clinical information. Volume data allows for a specific point in space to be evaluated from many different orienta tions by rotating, slicing, and referencing the slice to other orthogonal slices. It also allows for new volume-rendering displays that show depth, curvature, and surface images not available with conventional methods. The current limitations of image resolution, intuitive interfaces for obtaining and displaying optimal images, and technologic limitations for data storage and manipulation (including real-time three-dimensional ultrasound) will surely be overcome in the near future.

Detection of Jarcho-Levin syndrome at 12 weeks' gestation by nuchal translucency screening and three-dimensional ultrasound.

Jarcho-Levin syndrome (JLS) or spondylothoracic dysotosis causes early newborn death from respiratory insufficiency. Prenatal diagnosis is possible using fetal ultrasound, but requires a high level of suspicion and definitive diagnosis may be delayed well into the second trimester. We present a case in which a combination of three-dimensional ultrasound and measurement of nuchal translucency thickness allowed successful detection of recurrent JLS at 12 weeks' gestation.

Three-dimensional ultrasonography and assessment of the first-trimester fetus.

We used transvaginal three-dimensional ultrasonography to assess the first-trimester fetus and compared the findings with contemporaneous two-dimensional ultrasonographic studies. Multiplanar three-dimensional ultrasonography provided good visualization of fetal anatomy and allowed fetal measurement and assessment of nuchal translucency thickness. Three-dimensional ultrasonography required significantly less time to perform and to interpret than two-dimensional ultrasonography. Three-dimensional ultrasonography is an effective means of assessing the first-trimester fetus and offers potential advantages over two-dimensional ultrasonography.

Value of three-dimensional US for optimizing guidance for ablating focal liver tumors.

PURPOSE: To determine if three-dimensional ultrasound (3D US), by nature of its ability to simultaneously evaluate structures in three orthogonal planes and to study relationships of devices to tumor(s) and surrounding anatomic structures from any desired orientation, adds significant additional information to real-time 2D US used for placement of devices for ablation of focal liver tumors. MATERIALS AND METHODS: Sixteen patients underwent focal ablation of 23 liver tumors during two intraoperative cryoablation (CA) procedures, three intraoperative radiofrequency ablation (RFA) procedures, 11 percutaneous ethanol injections (PEI) procedures, and six percutaneous RFA procedures. After satisfactory placement of the ablative device(s) with 2D US guidance, 3D US was used to reevaluate adequacy to device position. Information added by 3D US and resultant alterations in device deployment were tabulated. RESULTS: 3D US added information in 20 of 22 (91%) procedures and caused the operator to readjust the number or position of ablative devices in 10 of 22 (45%) of procedures. Specifically, 3D US improved visualization and confident localization of devices in 13 of 22 (59%) procedures, detected unacceptable device placement in 10 of 22 (45%), and determined that 2D US had incorrectly predicted device orientation to a tumor in three of 22 (14%). CONCLUSIONS: Compared to conventional 2D US, 3D US provides additional relationship information for improved placement and optimal distribution of ablative agents for treatment of focal liver malignancy.