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A scarlet fever outbreak occurred in Hong Kong in 2011. The majority of cases resulted in the isolation of Streptococcus pyogenes emm12 with multiple antibiotic resistances. Phylogenetic analysis of 22 emm12 scarlet fever outbreak isolates, 7 temporally and geographically matched emm12 non-scarlet fever isolates, and 18 emm12 strains isolated during 2005-2010 indicated the outbreak was multiclonal. Genome sequencing of 2 nonclonal scarlet fever isolates (HKU16 and HKU30), coupled with diagnostic polymerase chain reaction assays, identified 2 mobile genetic elements distributed across the major lineages: a 64.9-kb integrative and conjugative element encoding tetracycline and macrolide resistance and a 46.4-kb prophage encoding superantigens SSA and SpeC and the DNase Spd1. Phenotypic comparison of HKU16 and HKU30 with the S. pyogenes M1T1 strain 5448 revealed that HKU16 displays increased adherence to HEp-2 human epithelial cells, whereas HKU16, HKU30, and 5448 exhibit equivalent resistance to neutrophils and virulence in a humanized plasminogen murine model. However, in contrast to M1T1, the virulence of HKU16 and HKU30 was not associated with covRS mutation. The multiclonal nature of the emm12 scarlet fever isolates suggests that factors such as mobile genetic elements, environmental factors, and host immune status may have contributed to the 2011 scarlet fever outbreak. [ABSTRACT FROM AUTHOR]

Competing Interests: All authors have disclosed no conflicts of interest.

The conventional medium chain chlorinated paraffin (MCCP) and zinc dialkyl dithiophosphate (ZDDP) additives have greatly enhanced the extreme pressure (EP) and anti-wear (AW) performance of the metalworking fluids. However, chlorine- and zinc-containing additives are restricted in use due to eco-toxicity issue. Herein, ashless and non-corrosive dimercaptobenzothiadiazole derivatives, namely bis-2,5-benzylsulfanyl-[1,3,4]thiadiazole (BBST) and bis-2,5-octylsulfanyl-[1,3,4]thiadiazole (BOST) consist of three sulfur atoms have been synthesized and evaluated. The performance of BBST shows a weld load ( P D ) of 3089 N and AW value of 5 mm 2 , which represents an improvement of 3.1 and 7.4 folds over naphthenic base oil (NBO). In addition, BBST also outperformed BOST, MCCP, and ZDDP in terms of its weld load and AW properties. Based on XPS analysis and molecular electrostatic potential maps (MEPS), BBST exhibits superior tribology performance due to the interaction between the sulfur (S), nitrogen (N), and π-electrons of the benzene ring with the metal surface. The formation of FeS, Fe 2 O 3 and Fe⋯N coordinate bonds contributes to the creation of an excellent tribofilm., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

A subcommittee of the Netherlands Commission on Radiation Dosimetry (NCS) was initiated in 2018 with the task to update and extend a previous publication (NCS-15) on the quality assurance of treatment planning systems (TPS) (Bruinvis et al 2005). The field of treatment planning has changed considerably since 2005. Whereas the focus of the previous report was more on the technical aspects of the TPS, the scope of this report is broader with a focus on a department wide implementation of the TPS. New sections about education, automated planning, information technology (IT) and updates are therefore added. Although the scope is photon therapy, large parts of this report will also apply to all other treatment modalities. This paper is a condensed version of these guidelines; the full version of the report in English is freely available from the NCS website (http://radiationdosimetry.org/ncs/publications). The paper starts with the scope of this report in relation to earlier reports on this subject. Next, general aspects of the commissioning process are addressed, like e.g. project management, education, and safety. It then focusses more on technical aspects such as beam commissioning and patient modeling, dose representation, dose calculation and (automated) plan optimisation. The final chapters deal with IT-related subjects and scripting, and the process of updating or upgrading the TPS., (© 2023 Institute of Physics and Engineering in Medicine.)

Background: Endometrial cancer is the most common gynecological cancer among women in developed countries. Sono-elastography is an extended ultrasonographic technique that has been shown to be useful in a wide range of conditions ranging from breast, prostate, and thyroid nodules to chronic liver disease and musculoskeletal conditions. The aim of this study is to compare the sonoelastographic features of endometrial malignancy and normal endometrium., Methods: This case-control observational study was conducted at a single institution. Participants with histologically proven endometrial cancer according to the results from microcurettage or hysteroscopic biopsy and scheduled for total hysterectomy were included as cases, while asymptomatic women scheduled for routine screening ultrasound examination were recruited as controls. Both cases and controls underwent conventional B-mode transvaginal ultrasonography and strain elastography. Demographic, ultrasonographic, and histopathologic findings were analyzed., Results: A total of 29 endometrial cancer patients (cases) and 28 normal females (controls) were included in the analysis. There was no significant difference in the mean age between the two groups, but the mean body weight was significantly higher in the case group ( P < 0.001). The strain ratio and elastographic thickness ratio of the endometrium were statistically significantly different between the case and the control group ( P ≤ 0.05) due to increased endometrial stiffness in cancer patients as compared to the normal group., Conclusion: Our results suggest that endometrial cancer can result in increased stiffness that is detectable by transvaginal sonoelastography. Sonoelastography may serve as an adjunct to conventional ultrasound in evaluating the endometrium of women with abnormal uterine bleeding., Competing Interests: Dr. Chiou Li Ong, an editorial board member at Journal of Medical Ultrasound, had no role in the peer review process of or decision to publish this article. The other authors declared no conflicts of interest in writing this paper., (Copyright: © 2022 Journal of Medical Ultrasound.)

This is a pictorial review on the radiological approach to patients with amenorrhea using a level-based framework. The prevalence of amenorrhea is 3 to 4% with wide-ranging causes involving multiple clinical disciplines. Normal menstruation depends on complex coordinated hormonal functions of the hypothalamic-pituitary-ovarian axis exerting its effect on an intact uterine end-organ and outflow tract. A disruption of any of these factors may result in amenorrhea. Categorizing the causes of primary and secondary amenorrhea into uterine, ovarian/gonadal, and intracranial levels provides a logical framework for its evaluation. A systematic level-based approach by targeted ultrasound of the pelvic structures is suggested, with different aims in primary versus secondary amenorrhea. Pelvic sonographic findings of various conditions within the uterine and ovarian/gonadal levels are illustrated. Conditions due to an intracranial cause result in downstream effects on the uterus and ovaries and can often be suspected based on a combination of clinical assessment, ultrasound findings, and laboratory investigations. By correlating pelvic ultrasound findings with underlying pathology, the clinical radiologist is able to provide useful diagnostic information in the management of these patients.

Intrauterine fetal demise (IUFD) is an uncommon but serious event that may occasionally be encountered on fetal MRI. Compared to the more florid signs of fetal demise which has occurred some time ago, recent IUFD is associated with more subtle findings that may be missed or misinterpreted. The two main MRI sequences used in imaging the fetus are T2-like two-dimensional balanced steady-state free-precession (SSFP), a white blood sequence, or T2-weighted single-shot fast spin-echo (SSFSE), a black blood sequence. The most reliable and specific signs of a recent IUFD are a constricted heart with poorly delineated cardiac chambers and signal abnormality in the heart and aorta, which will have different features depending on the MRI sequence used. Secondary signs of IUFD include global brain ischemia, abnormal globes, effusions, body wall edema and umbilical cord thrombosis. Unlike fetal ultrasound examinations where cardiac activity is routinely assessed, fetal MRI requires careful scrutiny of the fetal heart for assessment of fetal life.

Background: Immobilization devices are crucial to minimize patient positioning uncertainties in radiotherapy (RT) treatments. Accurate inter and intra-fraction motions is particularly important for intracranial and stereotactic radiation treatment which require high precision in dose delivery. Recently, a new immobilization device has been developed specifically for the radiation treatment of intracranial malignancies. To date, no data are available on the use of this device in daily clinical practice. The aim of this study is to investigate the intra and inter-fraction variations, patient comfort and radiographer confidence of the immobilization system from two distinct institutions: HagaZiekenhuis, Den Haag, Netherlands and IRCCS Ospedale Sacro Cuore Don Calabria, Negrar, Italy., Material and Method: Sixteen patients (10 diagnosed with brain metastases and 6 with primary central nervous systemic tumor) from IRCCS Ospedale Sacro Cuore Don Calabria and 17 patients (all diagnosed with brain metastases tumor) from HagaZiekenhuis were included in this study. The median target volume was 436 cc (range 3.2-1628 cc) and 4.58 cc (range 0.4-27.19 cc) for IRCCS and Haga, respectively. For patients treated in IRCCS Sacro Cuore Don Calabria, the median dose prescription was 30 Gy (range 27-60 Gy) and median number of fractions 10 (range 3-30). In Haga the median dose prescription was 21 Gy (range 8-21 Gy) and the median number of fraction was 1 (range 1-3). The immobilization device was assembled during CT simulation. A short interview to the patient regarding the device's comfort level was conducted at the end of the simulation procedure. Additionally, simulation setup time and radiographer (RTT) procedures (i.e. mask preparation) were evaluated. Prior to radiation treatment delivery, an automatic rigid match on the cranial bones between cone beam computed tomography (CBCT) and planning-CT was performed. A couch shift was performed subsequently. An extra post-treatment CBCT was acquire after the treatment delivery. This post-treatment CBCT was matched with pre-treatment CBCT to identify any possible intra-fraction motion. All online matches were validated by experienced radiation oncologist or RTT. A total of 126 CBCT's were analyzed offline by radiation oncologist/medical physicist. The data of the pre-treatment CBCT match was used to quantify inter-fraction motion. The post-treatment CBCT was matched with pre-treatment CBCT to identify any possible intra-fraction motion., Results: During the molding of the mask, all patients responded positive to the comfort. Median time required by the RTTs to assemble the immobilization system was 9 min (range 6-12 min). In terms of comfort, all patients reported a good-to high level of satisfaction. The RTTs also respond positively towards the use of the locking mechanism and clips. Results of positioning uncertainties were comparable between the two institutes. The mean inter-fraction motion for all translational and rotational directions were < 2 mm (SD < 4 mm) and < 0.5°(SD < 1.5°), respectively, while the mean intra-fraction motions were < 0.4 mm (SD < 0.6 mm) and 0.3° (SD < 0.5°)., Conclusions: This study demonstrates the efficacy and feasibility of the immobilization device in the intracranial radiation treatment. Both patient comfort and preparation time by RTTs are considered adequate. In combination with online daily imaging procedure, this device can achieve submillimeter accuracy required for intracranial and stereotactic treatments.

Introduction: Stereotactic radiosurgery (SRS) is a promising treatment option for patients with multiple brain metastases (BM). Recent technical advances have made LINAC based SRS a patient friendly technique, allowing for accurate patient positioning and a short treatment time. Since SRS is increasingly being used for patients with multiple BM, it remains essential that SRS be performed with the highest achievable quality in order to prevent unnecessary complications such as radionecrosis. The purpose of this article is to provide guidance for high-quality LINAC based SRS for patients with BM, with a focus on single isocenter non-coplanar volumetric modulated arc therapy (VMAT). Methods: The article is based on a consensus statement by the study coordinators and medical physicists of four trials which investigated whether patients with multiple BM are better palliated with SRS instead of whole brain radiotherapy (WBRT): A European trial (NCT02353000), two American trials and a Canadian CCTG lead intergroup trial (CE.7). This manuscript summarizes the quality assurance measures concerning imaging, planning and delivery. Results: To optimize the treatment, the interval between the planning-MRI (gadolinium contrast-enhanced, maximum slice thickness of 1.5 mm) and treatment should be kept as short as possible (< two weeks). The BM are contoured based on the planning-MRI, fused with the planning-CT. GTV-PTV margins are minimized or even avoided when possible. To maximize efficiency, the preferable technique is single isocenter (non-)coplanar VMAT, which delivers high doses to the target with maximal sparing of the organs at risk. The use of flattening filter free photon beams ensures a lower peripheral dose and shortens the treatment time. To bench mark SRS treatment plan quality, it is advisable to compare treatment plans between hospitals. Conclusion: This paper provides guidance for quality assurance and optimization of treatment delivery for LINAC-based radiosurgery for patients with multiple BM.

Coarctation of the aorta (CoA) is present in 0.4% of live births and in 7% of patients with congenital heart disease. While there may be florid presentations of congestive heart failure in the neonatal period, the diagnosis during adulthood is often delayed. We encountered a 20-year-old woman who was discovered to be hypertensive on routine check-up. Following bilateral abnormal renal doppler sonography, MR angiogram revealed a short-segment stenosis of the descending thoracic aorta. Review of her chest radiograph showed a small aortic knuckle. This case highlights an unconventional algorithm in diagnosing aortic coarctation in adulthood.

Background and Purpose: High fidelity human patient simulation (HPS) has been incorporated in various United States doctor of pharmacy programs with favorable learning experiences, knowledge retention, and problem-solving skills reported. In Singapore, HPS is a novel learning technique as it has not been utilized in the Bachelor of Science (Pharmacy) curriculum or for continuing professional education (CPE). It is necessary to evaluate acceptance of HPS compared to asynchronous online learning (AOL)., Educational Activity and Setting: Nineteen participants from two institutions completed the study in Singapore. This was an experimental study design with participants randomized into groups A and B. Group A completed AOL followed by HPS, whereas group B completed them in reverse order. Acceptance of teaching modalities was evaluated with a Likert scale survey and analyzed with Fisher's exact test. Educational content was congestive heart failure and was evaluated externally for equivalency., Findings: All participants enjoyed the HPS activity compared to 13 (68.4%; p = 0.02) for AOL. Eighteen (94.7%) and 15 (78.9%; p = 0.34) participants felt that HPS activity and AOL improved their critical and decision-making skills respectively. Sixteen (84.2%) and 17 (89.5%) agreed that AOL and HPS activity improved their confidence (p = 1.00)., Discussion: Participants enjoyed HPS activity significantly more than AOL. HPS activity could be used to achieve improved critical and decision-making skills of learners as there was a trend of more learners perceiving improvement compare to AOL., Summary: High fidelity HPS learning was well received by participants in Singapore and can be implemented in CPE., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Antral follicle count (AFC) and anti-Mullerian hormone (AMH) are known as the most reliable markers of a woman's ovarian reserve and are related to age. There is currently no specific local age-related centile charts for AFC and AMH. Therefore, we aim to examine the relationship between AFC and AMH with age and construct age-related nomograms among a subfertile Asian population., Methods: This is a study involving Chinese women who had their AFC and AMH measured as part of their subfertility screening from December 2010 until November 2014 in KK Women's and Children's Hospital, Singapore. Ordinary least squares regression analysis was used to estimate the relationship of AFC and AMH with age, while age-related AFC and AMH nomograms for the 3rd, 10th, 25th, 50th, 75th, 90th and 97th percentiles were produced using the lambda-mu-sigma method., Results: A total of 1,009 women, aged 26 to 44 year-old, were included. On average, the AFC and AMH decreased respectively by 0.79 follicle (95% confidence interval -0.93, -0.64) and 0.38 ng/mL (95% confidence interval -0.43, -0.32) per year of age. The age-related nomograms of AFC showed an approximately linear pattern, inversely correlated with age, regardless of the percentile. For AMH, the pattern is linear for the 75th percentile and below but shows a slightly accelerating decline for the 90th and 97th percentile. Overall, there were large inter-individual variations in AFC and AMH up to about 40 year-old., Conclusion: The declines of AFC and AMH over age are mostly linear among subfertile Chinese women in Singapore. The age-related AFC and AMH nomograms could be used as a reference chart by fertility practitioners. However, future validation with longitudinal data is required.

We aimed to characterize antimicrobial zinc trafficking within macrophages and to determine whether the professional intramacrophage pathogen Salmonella enterica serovar Typhimurium (S Typhimurium) subverts this pathway. Using both Escherichia coli and S Typhimurium, we show that TLR signaling promotes the accumulation of vesicular zinc within primary human macrophages. Vesicular zinc is delivered to E. coli to promote microbial clearance, whereas S. Typhimurium evades this response via Salmonella pathogenicity island (SPI)-1. Even in the absence of SPI-1 and the zinc exporter ZntA, S Typhimurium resists the innate immune zinc stress response, implying the existence of additional host subversion mechanisms. We also demonstrate the combinatorial antimicrobial effects of zinc and copper, a pathway that S. Typhimurium again evades. Our use of complementary tools and approaches, including confocal microscopy, direct assessment of intramacrophage bacterial zinc stress responses, specific E. coli and S Typhimurium mutants, and inductively coupled plasma mass spectroscopy, has enabled carefully controlled characterization of this novel innate immune antimicrobial pathway. In summary, our study provides new insights at the cellular level into the well-documented effects of zinc in promoting host defense against infectious disease, as well as the complex host subversion strategies employed by S Typhimurium to combat this pathway.-Kapetanovic, R., Bokil, N. J., Achard, M. E. S., Ong, C.-L. Y., Peters, K. M., Stocks, C. J., Phan, M.-D., Monteleone, M., Schroder, K., Irvine, K. M., Saunders, B. M., Walker, M. J., Stacey, K. J., McEwan, A. G., Schembri, M. A., Sweet, M. J. Salmonella employs multiple mechanisms to subvert the TLR-inducible zinc-mediated antimicrobial response of human macrophages., (© FASEB.)

Methylglyoxal is a dicarbonyl compound that acts as a toxic electrophile in biological systems. Methylglyoxal is produced in certain bacteria as a byproduct of glycolysis through methylglyoxal synthase. Like many bacteria, Group A Streptococcus (GAS), a Gram-positive human pathogen responsible for a wide spectrum of diseases, uses a two-step glyoxalase system to remove methylglyoxal. However, bioinformatic analysis revealed that no homologue of methylglyoxal synthase is present in GAS, suggesting that the role of the glyoxalase system is to detoxify methylglyoxal produced by the host. In this study, we investigated the role of methylglyoxal detoxification in the pathogenesis of GAS. A mutant (5448ΔgloA), deficient in glyoxylase I (S-lactoylglutathione lyase), was constructed and tested for susceptibility to methylglyoxal, human neutrophil survival and virulence in a murine model of infection. 5448ΔgloA was more sensitive to methylglyoxal and was also more susceptible to human neutrophil killing. Inhibition of neutrophil myeloperoxidase rescued the gloA-deficient mutant indicating that this enzyme was required for methylglyoxal production. Furthermore, the 5448ΔgloA mutant was slower at disseminating into the blood in the murine model. These data suggest that neutrophils produce methylglyoxal as an antimicrobial agent during bacterial infection, and the glyoxalase system is part of the GAS defence against the innate immune system during pathogenesis., (© FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Ultrasonography (US) is the most recent cross-sectional imaging modality to acquire three-dimensional (3D) capabilities. The reconstruction of volumetric US data for multiplanar display took a significantly longer time to develop in comparison with computed tomography and magnetic resonance imaging. The current equipment for 3D-US is capable of producing high-resolution images in three different planes, including real-time surface-rendered images. The use of 3D-US in gynaecology was accelerated through the development of the endovaginal volume transducer, which allows the automated acquisition of volumetric US data. Although initially considered an adjunct to two-dimensional US, 3D-US is now the imaging modality of choice for the assessment of Müllerian duct anomalies and the location of intrauterine devices.

The group A Streptococcus (GAS) M1T1 clone emerged in the 1980s as a leading cause of epidemic invasive infections worldwide, including necrotizing fasciitis and toxic shock syndrome. Horizontal transfer of mobile genetic elements has played a central role in the evolution of the M1T1 clone, with bacteriophage-encoded determinants DNase Sda1 and superantigen SpeA2 contributing to enhanced virulence and colonization respectively. Outbreaks of scarlet fever in Hong Kong and China in 2011, caused primarily by emm12 GAS, led to our investigation of the next most common cause of scarlet fever, emm1 GAS. Genomic analysis of 18 emm1 isolates from Hong Kong and 16 emm1 isolates from mainland China revealed the presence of mobile genetic elements associated with the expansion of emm12 scarlet fever clones in the M1T1 genomic background. These mobile genetic elements confer expression of superantigens SSA and SpeC, and resistance to tetracycline, erythromycin and clindamycin. Horizontal transfer of mobile DNA conferring multi-drug resistance and expression of a new superantigen repertoire in the M1T1 clone should trigger heightened public health awareness for the global dissemination of these genetic elements.

Isolated fallopian tube torsion is an uncommon diagnosis. It is particularly rare in the paediatric and post-menopausal age groups. It lacks pathognomonic symptoms, signs and imaging findings, yet each of these diagnostic steps plays a crucial role in early diagnosis. We describe two cases of isolated fallopian tube torsion in prepubertal females.

Zinc (Zn) and copper (Cu) are essential for optimal innate immune function, and nutritional deficiency in either metal leads to increased susceptibility to bacterial infection. Recently, the decreased survival of bacterial pathogens with impaired Cu and/or Zn detoxification systems in phagocytes and animal models of infection has been reported. Consequently, a model has emerged in which the host utilizes Cu and/or Zn intoxication to reduce the intracellular survival of pathogens. This review describes and assesses the potential role for Cu and Zn intoxication in innate immune function and their direct bactericidal function., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)

Neutrophils release free zinc to eliminate the phagocytosed bacterial pathogen Streptococcus pyogenes (Group A Streptococcus; GAS). In this study, we investigated the mechanisms underpinning zinc toxicity towards this human pathogen, responsible for diseases ranging from pharyngitis and impetigo, to severe invasive infections. Using the globally-disseminated M1T1 GAS strain, we demonstrate that zinc stress impairs glucose metabolism through the inhibition of the glycolytic enzymes phosphofructokinase and glyceraldehyde-3-phosphate dehydrogenase. In the presence of zinc, a metabolic shift to the tagatose-6-phosphate pathway allows conversion of D-galactose to dihydroxyacetone phosphate and glyceraldehyde phosphate, partially bypassing impaired glycolytic enzymes to generate pyruvate. Additionally, zinc inhibition of phosphoglucomutase results in decreased capsule biosynthesis. These data indicate that zinc exerts it toxicity via mechanisms that inhibit both GAS central carbon metabolism and virulence pathways.

Unlabelled: Streptococcus pyogenes (group A Streptococcus [GAS]) is an obligate human pathogen responsible for a spectrum of human disease states. Metallobiology of human pathogens is revealing the fundamental role of metals in both nutritional immunity leading to pathogen starvation and metal poisoning of pathogens by innate immune cells. Spy0980 (MntE) is a paralog of the GAS zinc efflux pump CzcD. Through use of an isogenic mntE deletion mutant in the GAS serotype M1T1 strain 5448, we have elucidated that MntE is a manganese-specific efflux pump required for GAS virulence. The 5448ΔmntE mutant had significantly lower survival following infection of human neutrophils than did the 5448 wild type and the complemented mutant (5448ΔmntE::mntE). Manganese homeostasis may provide protection against oxidative stress, explaining the observed ex vivo reduction in virulence. In the presence of manganese and hydrogen peroxide, 5448ΔmntE mutant exhibits significantly lower survival than wild-type 5448 and the complemented mutant. We hypothesize that MntE, by maintaining homeostatic control of cytoplasmic manganese, ensures that the peroxide response repressor PerR is optimally poised to respond to hydrogen peroxide stress. Creation of a 5448ΔmntE-ΔperR double mutant rescued the oxidative stress resistance of the double mutant to wild-type levels in the presence of manganese and hydrogen peroxide. This work elucidates the mechanism for manganese toxicity within GAS and the crucial role of manganese homeostasis in maintaining GAS virulence., Importance: Manganese is traditionally viewed as a beneficial metal ion to bacteria, and it is also established that most bacteria can tolerate high concentrations of this transition metal. In this work, we show that in group A Streptococcus, mutation of the mntE locus, which encodes a transport protein of the cation diffusion facilitator (CDF) family, results in accumulation of manganese and sensitivity to this transition metal ion. The toxicity of manganese is indirect and is the result of a failure of the PerR regulator to respond to oxidative stress in the presence of high intracellular manganese concentrations. These results highlight the importance of MntE in manganese homeostasis and maintenance of an optimal manganese/iron ratio in GAS and the impact of manganese on resistance to oxidative stress and virulence., (Copyright © 2015 Turner et al.)

Cadmium is a transition metal ion that is highly toxic in biological systems. Although relatively rare in the Earth's crust, anthropogenic release of cadmium since industrialization has increased biogeochemical cycling and the abundance of the ion in the biosphere. Despite this, the molecular basis of its toxicity remains unclear. Here we combine metal-accumulation assays, high-resolution structural data and biochemical analyses to show that cadmium toxicity, in Streptococcus pneumoniae, occurs via perturbation of first row transition metal ion homeostasis. We show that cadmium uptake reduces the millimolar cellular accumulation of manganese and zinc, and thereby increases sensitivity to oxidative stress. Despite this, high cellular concentrations of cadmium (~17 mM) are tolerated, with negligible impact on growth or sensitivity to oxidative stress, when manganese and glutathione are abundant. Collectively, this work provides insight into the molecular basis of cadmium toxicity in prokaryotes, and the connection between cadmium accumulation and oxidative stress.

Introduction: Uterine fibroids are the most common type of gynaecologic benign tumours, occurring in 25% to 50% of women during their reproductive lives. About half of the affected women have clinically significant symptoms, including abnormal bleeding, menstrual pain, frequent urination, constipation and abdominal distension. Magnetic resonance-guided focused ultrasound surgery (MRgFUS) has been used to treat patients with benign lesions and a variety of malignancies. The objective of this study is to evaluate symptom relief before and after MR-guided ultrasound ablation of fibroids., Materials and Methods: A total of 37 patients with symptomatic uterine fibroids were treated in this study., Results: MRgFUS treatment led to a significant, time-dependent decrease in not only Symptom Severity Scores (SSS), but also the mean fibroid volume. The average reductions in volume were 41.6% and 52.6% at 6 months and 12 months respectively (P <0.05). The mean SSS of the 37 patients was 41.7 ± 2.8 before treatment whereas the average SSS was 26.9 ± 3.6, 20.7 ± 3.4, 18.5 ± 3.6, 16.5 ± 7.1, 9.8 ± 3.6 at 3 months, 6 months, 1 year, 2 years, and 3 to 4 years respectively. The decrease in scores was significant at all time points up to 3 to 4 years (P <0.05 and P <0.001)., Conclusion: MRgFUS is a safe and effective non-invasive treatment for patients with symptomatic fibroids.

Lactate is an abundant metabolite, produced by host tissues and commensal organisms, and it represents an important potential carbon source for bacterial pathogens. In the case of Neisseria spp., the importance of the lactate permease in colonization of the host has been demonstrated, but there have been few studies of lactate metabolism in pathogenic Neisseria in the postgenomic era. We describe herein the characterization of genome-annotated, respiratory, and substrate-level lactate dehydrogenases (LDHs) from the obligate human pathogen Neisseria gonorrhoeae. Biochemical assays using N. gonorrhoeae 1291 wild type and isogenic mutant strains showed that cytoplasmic LdhA (NAD(+)-dependent D-lactate dehydrogenase) and the membrane-bound respiratory enzymes, LdhD (D-lactate dehydrogenase) and LldD (L-lactate dehydrogenase) are correctly annotated. Mutants lacking LdhA and LdhD showed greatly reduced survival in neutrophils compared with wild type cells, highlighting the importance of D-lactate metabolism in gonococcal survival. Furthermore, an assay of host colonization using the well-established human primary cervical epithelial cell model revealed that the two respiratory enzymes make a significant contribution to colonization of and survival within the microaerobic environment of the host. Taken together, these data suggest that host-derived lactate is critical for the growth and survival of N. gonorrhoeae in human cells., (© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Background: Zinc plays an important role in human immunity, and it is known that zinc deficiency in the host is linked to increased susceptibility to bacterial infection. In this study, we investigate the role of zinc efflux in the pathogenesis of Streptococcus pyogenes (group A Streptococcus [GAS]), a human pathogen responsible for superficial infections, such as pharyngitis and impetigo, and severe invasive infections., Methods: The clinically important M1T1 wild-type strain was used in this study, and isogenic mutants were constructed with deletions in the czcD gene (Spy0653; which encodes a putative zinc efflux pump) and adjacent gczA gene (Spy0654; which encodes a putative zinc-dependent activator of czcD). Wild-type, isogenic mutants and complemented strains were tested for resistance against zinc stress, intracellular zinc accumulation, and virulence., Results: Both czcD and gczA mutants exhibited increased sensitivity to zinc. Transcriptional analyses indicate that GczA upregulates czcD in response to zinc. Both mutants displayed increased susceptibility to human neutrophil killing and reduced virulence in a murine infection model. Furthermore, we showed that neutrophils mobilize zinc in response to GAS., Conclusions: These data indicate that the innate immune system may use zinc as an antimicrobial agent and that zinc efflux is an important contributor to GAS pathogenesis.

Streptococcus pneumoniae requires manganese for colonization of the human host, but the underlying molecular basis for this requirement has not been elucidated. Recently, it was shown that zinc could compromise manganese uptake and that zinc levels increased during infection by S. pneumoniae in all the niches that it colonized. Here we show, by quantitative means, that extracellular zinc acts in a dose dependent manner to competitively inhibit manganese uptake by S. pneumoniae, with an EC50 of 30.2 µM for zinc in cation-defined media. By exploiting the ability to directly manipulate S. pneumoniae accumulation of manganese, we analyzed the connection between manganese and superoxide dismutase (SodA), a primary source of protection for S. pneumoniae against oxidative stress. We show that manganese starvation led to a decrease in sodA transcription indicating that expression of sodA was regulated through an unknown manganese responsive pathway. Intriguingly, examination of recombinant SodA revealed that the enzyme was potentially a cambialistic superoxide dismutase with an iron/manganese cofactor. SodA was also shown to provide the majority of protection against oxidative stress as a S. pneumoniae ΔsodA mutant strain was found to be hypersensitive to oxidative stress, despite having wild-type manganese levels, indicating that the metal ion alone was not sufficiently protective. Collectively, these results provide a quantitative assessment of the competitive effect of zinc upon manganese uptake and provide a molecular basis for how extracellular zinc exerts a 'toxic' effect on bacterial pathogens, such as S. pneumoniae.

Ectopic pregnancy in the interstitial part of the fallopian tube is a rare event, associated with a high rate of complications due to delayed diagnosis. Rupture of such pregnancy often results in catastrophic hemorrhage. Several reports highlighted the role of magnetic resonance imaging in establishing the diagnosis of interstitial pregnancy, but magnetic resonance imaging findings of a ruptured advanced interstitial ectopic pregnancy have not been published before. The authors therefore present characteristic findings on magnetic resonance imaging of a ruptured interstitial ectopic pregnancy which had reached 18 weeks, in a 25-year-old woman who presented with acute abdominal pain.

Purpose: We investigated the dosimetric impact of the interplay effect during RapidArc stereotactic body radiation therapy for lung tumors using flattening filter-free (FFF) beams with different dose rates., Methods and Materials: Seven tumors with motion ≤20 mm, treated with 10-MV FFF RapidArc, were analyzed. A programmable phantom with sinusoidal longitudinal motion (30-mm diameter "tumor" insert; period = 5 s; individualized amplitude from planning 4-dimensional computed tomography) was used for dynamic dose measurements. Measurements were made with GafChromic EBT III films. Plans delivered the prescribed dose to 95% of the planning target volume, created by a 5-mm expansion of the internal target volume. They comprised 2 arcs and maximum dose rates of 400 and 2400 MU/min. For 2400 MU/min plans, measurements were repeated at 3 different initial breathing phases to model interplay over 2 to 3 fractions. For 3 cases, 2 extra plans were created using 1 full rotational arc (with contralateral lung avoidance sector) and 1 partial arc of 224° to 244°. Dynamic and convolved static measurements were compared by use of gamma analysis of 3% dose difference and 1 mm distance-to-agreement., Results: For 2-arc 2400 MU/min plans, maximum dose deviation of 9.4% was found in a single arc; 7.4% for 2 arcs (single fraction) and <5% and 3% when measurements made at 2 and 3 different initial breathing phases were combined, simulating 2 or 3 fractions. For all 7 cases, >99% of the area within the region of interest passed the gamma criteria when all 3 measurements with different initial phases were combined. Single-fraction single-arc plans showed higher dose deviations, which diminished when dose distributions were summed over 2 fractions. All 400 MU/min plans showed good agreement in a single fraction measurement., Conclusion: Under phantom conditions, single-arc and single-fraction 2400 MU/min FFF RapidArc lung stereotactic body radiation therapy is susceptible to interplay. Two arcs and ≥2 fractions reduced the effect to a level that appeared unlikely to be clinically significant., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Purpose: To study the dosimetric impact of relatively short-duration intrafraction shifts during a single fraction of RapidArc delivery for vertebral stereotactic body radiation therapy (SBRT) using flattened (FF) and flattening filter-free (FFF) beams., Methods and Materials: The RapidArc plans, each with 2 to 3 arcs, were generated for 9 patients using 6-MV FF and 10-MV FFF beams with maximum dose rates of 1000 and 2400 MU/min, respectively. A total of 1272 plans were created to estimate the dosimetric consequences in target and spinal cord volumes caused by intrafraction shifts during one of the arcs. Shifts of 1, 2, and 3 mm for periods of 5, 10, and 30 seconds, and 5 mm for 5 and 10 seconds, were modelled during a part of the arc associated with high doses and steep dose gradients., Results: For FFF plans, shifts of 2 mm over 10 seconds and 30 seconds could increase spinal cord Dmax by up to 6.5% and 13%, respectively. Dosimetric deviations in FFF plans were approximately 2-fold greater than in FF plans. Reduction in target coverage was <1% for 83% and 96% of the FFF and FF plans, respectively., Conclusion: Even short-duration intrafraction shifts can cause significant dosimetric deviations during vertebral SBRT delivery, especially when using very high dose rate FFF beams and when the shift occurs in that part of the arc delivering high doses and steep gradients. The impact is greatest on the spinal cord and its planning-at-risk volume. Accurate and stable patient positioning is therefore required for vertebral SBRT., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The resurgence of invasive disease caused by Streptococcus pyogenes (group A Streptococcus [GAS]) in the past 30 years has paralleled the emergence and global dissemination of the highly virulent M1T1 clone. The GAS M1T1 clone has diverged from the ancestral M1 serotype by horizontal acquisition of two unique bacteriophages, encoding the potent DNase Sda1/SdaD2 and the superantigen SpeA, respectively. The phage-encoded DNase promotes escape from neutrophil extracellular traps and is linked to enhanced virulence of the M1T1 clone. In this study, we successfully used in vitro lysogenic conversion to transfer the Sda1-encoding phage from the M1T1 clonal strain 5448 to the nonclonal M1 isolate SF370 and determined the impact of this horizontal gene transfer event on virulence. Although Sda1 was expressed in SF370 lysogens, no capacity of the phage-converted strain to survive human neutrophil killing, switch to a hyperinvasive covRS mutant form, or cause invasive lethal infection in a humanized plasminogen mouse model was observed. This work suggests that the hypervirulence of the M1T1 clone is due to the unique synergic effect of the M1T1 clone bacteriophage-specific virulence factor Sda1 acting in concert with the M1T1 clone-specific genetic scaffold.

Streptococcus pneumoniae (the pneumococcus) is a major human pathogen that is carried asymptomatically in the nasopharynx by up to 70% of the human population. Translocation of the bacteria into internal sites can cause a range of diseases, such as pneumonia, otitis media, meningitis, and bacteremia. This transition from nasopharynx to growth at systemic sites means that the pneumococcus needs to adjust to a variety of environmental conditions, including transition metal ion availability. Although it is an important nutrient, iron potentiates oxidative stress, and it is established that in S. pneumoniae, expression of iron transport systems and proteins that protect against oxidative stress are regulated by an orphan response regulator, RitR. In this study, we investigated the effect of iron and manganese ion availability on the growth of a ritR mutant. Deletion of ritR led to impaired growth of bacteria in high-iron medium, but this phenotype could be suppressed with the addition of manganese. Measurement of metal ion accumulation indicated that manganese prevents iron accumulation. Furthermore, the addition of manganese also led to a reduction in the amount of hydrogen peroxide produced by bacterial cells. Studies of virulence in a murine model of infection indicated that RitR was not essential for pneumococcal survival and suggested that derepression of iron uptake systems may enhance the survival of pneumococci in some niches.

Purpose: Flattening filter-free (FFF) beams with higher dose rates and faster delivery are now clinically available. The purpose of this planning study was to compare optimized non-FFF and FFF RapidArc plans for stereotactic body radiotherapy (SBRT) and to validate the accuracy of fast arc delivery., Methods and Material: Ten patients with peripheral lung tumors and 10 with vertebral metastases were planned using RapidArc with a flattened 6-MV photon beam and a 10-MV FFF beam for fraction doses of 7.5-18 Gy. Dosimetry of the target and organs at risk (OAR), number of monitor units (MU), and beam delivery times were assessed. GafChromic EBT2 film measurements of FFF plans were performed to compare calculated and delivered dose distributions., Results: No major dosimetric differences were seen between the two delivery techniques. For lung SBRT plans, conformity indices and OAR doses were similar, although the average MU required were higher with FFF plans. For vertebral SBRT, FFF plans provided comparable PTV coverage, with no significant differences in OAR doses. Average beam delivery times were reduced by a factor of up to 2.5, with all FFF fractions deliverable within 4 min. Measured FFF plans showed high agreement with calculated plans, with more than 99% of the area within the region of interest fulfilling the acceptance criterion., Conclusion: The higher dose rate of FFF RapidArc reduces delivery times significantly, without compromising plan quality or accuracy of dose delivery., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: This study aimed to develop and provisionally validate a novel scoring index for preoperative cancer-risk prediction in childhood ovarian tumors., Methods: Fifty-five girls aged 18 and below underwent surgery for ovarian masses between 2004 and 2009. Benign or non-benign histological diagnoses (the latter including all malignant and borderline tumors and tumors containing immature components) were correlated with clinical and biochemical parameters, and blinded scores of ultrasound and computed-tomography using multivariate logistic regression. Regression coefficients were used as weighting factors to create an additive index. This index was validated prospectively against 23 consecutive adnexal masses operated in 2010., Results: In total, 67 tumors were benign and 11 non-benign. Non-benign diagnosis was independently associated with the maximum diameter of the largest solid component (score = value in cm), the presence of sex hormone-related symptoms (score = +6), and enhancement or flow in a septum or solid papillary projection (score = +4). The novel scoring index was calculated as the total score of these three parameters. A cutoff score of 7 gave a specificity of 97.9% and sensitivity of 87.5% for the training data set, and specificity and sensitivity of 100% for the pilot testing set., Conclusion: The novel pediatric risk-of-malignancy index is able to accurately discriminate between benign and non-benign ovarian tumors in children and adolescents. Its preoperative application may guide surgical management decisions before the availability of histological confirmation.

This study is a retrospective review of the transcervical fluoroscopy-guided fallopian tube recanalisation (FTR) procedures done in a multi-ethnic south-east Asian population, over 9 years. A total of 100 patients with infertility and documented proximal tubal obstruction (PTO) were referred for FTR. On-table hysterosalpingography under sedation demonstrated true PTO in 96 patients. At selective-salpingography, the PTO cleared in 16 patients; 78 required FTR and two had fimbrial blockage. The technical success rate of FTR was 86.8% and the post-FTR pregnancy rate was 36.84% at a mean follow-up interval of 12.2 months. There were no major, immediate procedure-related complications. There was an ectopic pregnancy in a single treated patient. Fluoroscopy-guided FTR is a safe treatment option in patients with infertility from PTO, with high technical success rate, low complication rate and increased chances of pregnancy; therefore it should be preferred before attempting more expensive and resource-intensive procedures.

Purpose: To investigate the impact of the calculation resolution of the anisotropic analytical algorithms (AAA) for a variety of small fields in homogeneous and heterogeneous media and for RapidArc plans., Methods: Dose distributions calculated using AAA version 8.6.15 (AAA8) and 10.0.25 (AAA10) were compared to measurements performed with GafChromic EBT film, using phantoms made of polystyrene or a combination of polystyrene and cork. The accuracy of the algorithms calculated using grid resolutions of 2.5 and 1.0 mm was investigated for different field sizes, and for a limited selection of RapidArc plans (head and neck, small meningioma, and lung). Additional plans were optimized to create excessive multileaf collimator modulation and measured on a homogenous phantom. Gamma evaluation criterion of 3% dose difference and 2- or 1-mm distance to agreement (DTA) were applied to evaluate the accuracy of the algorithms., Results: For fields < or = 3 x 3 cm2, both versions of AAA predicted lower peak doses and broader penumbra widths than the measurements. However, AAA10 and a finer calculation grid improved the agreement. For RapidArc plans with many small multileaf collimator (MLC) segments and relatively high number of monitor units (MU), AAA8 failed to identify small dose peaks within the target. Both versions performed better in polystyrene than in cork. In homogeneous cork layers, AAA8 underestimated the average target dose for a clinical lung plan. This was improved with AAA10 calculated using a 1 mm grid., Conclusions: AAA10 improves the accuracy of dose calculations, and calculation grid of 1.0 mm is superior to using 2.5 mm, although calculation times increased by factor of 5. A suitable upper MU constraint should be assigned during optimization to avoid plans with high modulation. For plans with a relative high number of monitor units, calculations using 1 mm grid resolution are recommended. For planning target volume (PTV) which contains relatively large area of low density tissue, users should be aware of possible dose underestimation in the low density region and recalculation with AAA10 grid 1.0 mm is recommended.

Introduction: The purpose of this case series was to determine the sonographic features of neonatal ovarian torsion., Materials and Methods: Seven surgically proven cases of neonatal ovarian cysts were included in this retrospective study. The patients were divided into 2 groups, torsion and non-torsion. These 7 patients were evaluated for the clinical presentation, sonographic features, surgical and pathological findings. The findings on follow-up sonography after surgery were also noted., Results: The sonographic appearance was variable. Of the 4 cases with torsion, 2 lesions had internal echoes with 'fi sh-net appearance'. The other 2 lesions were predominantly cystic on the sonography with internal echoes and echogenic nodule. A calcific focus was present in 1 of these echogenic nodules. One of the cysts had fluid-fluid level. In the non-torsion group, only 1 lesion had mixed echogenic appearance. The other 2 lesions were cystic with low level internal echoes in 1 of the cysts. The surgical procedure performed in the torsion group was salpingo-oophorectomy in 2 patients and oophorectomy in 1 patient. In 1 patient, cystectomy was attempted without success. In the non-torsion group, only cystectomy was performed with preservation of normal ovaries, which was confirmed on follow-up sonography., Conclusion: The sonographic features of cysts with 'fish-net appearance', fluid-debris level and cysts with echogenic nodule favour torsion. The former sign has so far not been described as a sonographic predictor for neonatal ovarian torsion.

Large extraluminal gastrointestinal tumors (GISTs) may present as pelvic masses and thus mimic gynecological neoplasms in female patients. On clinical examination and pelvic ultrasound, these tumors resemble pedunculated fibroids or ovarian tumors. Multidetector computed tomography (CT), with its ability to perform isotropic multiplanar reconstruction, is useful in differentiating GISTs from true gynecological masses by demonstrating the pedicle sign connecting a pelvic GIST to its organ of origin. This allows a preoperative diagnosis to be made, which may be helpful in guiding therapeutic options and management. We present two cases of GISTs presenting as pelvic masses in which ultrasound findings suggested a gynecological cause, but multidetector CT with multiplanar reconstruction was able to determine their true organ of origin.

Purpose: To study the dosimetric predictors of early clinical toxicity following SBRT in patients with lung tumors and planning target volumes (PTV) exceeding 80 cm(3)., Methods: Eighteen consecutive patients who were treated using volumetric modulated arc therapy (RapidArc™) were assessed. All were either unfit or refused to undergo surgery or chemoradiotherapy. PTV planning objectives were as used in the ROSEL study protocol. Clinical toxicity was scored using Common Toxicity Criteria AE4.0. Lung volumes receiving 5, 10, 15, and 20 Gy (V(5), V(10), V(15) and V(20)) and mean lung dose were assessed and correlated to symptomatic radiation pneumonitis (RP)., Results: Median age, age-adjusted Charlson-comorbidity score and PTV size were 74, 7.5 and 137 cm(3), respectively. At a median follow-up of 12.8 months, 8 deaths were recorded: 5 arising from comorbidity, 2 were potentially treatment-related and 1 had local recurrence. RP was reported in 5 patients (grade 2 in 3 and grade 3 in 2). All RP occurred in plans without a high priority optimization objective on contralateral lung. Acute RP was best predicted by contralateral lung V(5) (p<0.0001)., Conclusion: After SBRT using RapidArc in lung tumors >80 cm(3), the contralateral lung V(5) best predicts RP. Limiting contralateral lung V(5) to <26% may reduce acute toxicity., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Purpose: Our aim was to present our experience with the management of six women with uterine scar pregnancies in KK Women's and Children's Hospital, Singapore., Methods: The medical records of women with a pregnancy in previous uterine scar that had been diagnosed in our department during 2004-2008 were reviewed., Results: Out of six women, one woman presented in mid-trimester, at 16 weeks with severe abdominal pain and persistent vomiting. She underwent a hysterectomy complicated with massive haemorrhage. The other five women presented in first trimester. Two women had excision of the scar with the sac, two had ultrasound-guided injection of methotrexate in the sac and one had systemic methotrexate. In all cases, maternal recovery was complete. Uterine scar pregnancy was diagnosed by ultrasonography., Conclusion: Women at a risk appear to be those with multiple Caesarean sections, termination of pregnancy and myomectomy. Operative as well as medical treatments have been reported for scar pregnancy. Surgical treatment includes excision of trophoblastic tissues by laparotomy or laparoscopy whilst medical treatment includes local and/or systemically administered methotrexate. Although many interventions have been described, optimal treatment is still not known and they remain a challenge.

Purpose: Volumetric modulated arc therapy (RapidArc) allows for fast delivery of stereotactic body radiotherapy (SBRT) delivery in stage I lung tumors. We compared dose distributions and delivery times between RapidArc and common delivery techniques in small tumors., Methods: In 18 patients who completed RapidArc SBRT for tumors measuring <70 cm(3), new treatment plans were generated using non-coplanar 3D conformal fields (conf-SBRT) and dynamic conformal arc radiotherapy (DCA). For 9 patients with tumors adjacent to the chest wall, co-planar intensity-modulated radiotherapy (IMRT) plans were also generated. PTV dose coverage, organs at risk (OAR) doses and treatment delivery times were assessed., Results: RapidArc plans achieved a superior conformity index (CI) and lower V(45 Gy) to chest wall (p<0.05) compared to all other techniques. RapidArc led to a small increase in V(5 Gy) to contralateral lung compared to conf-SBRT (4.4±4% versus 1.2±1.8%, p=0.011). For other OAR, RapidArc and conf-SBRT plans were comparable, and both were superior to DCA plans. Delivery of a 7.5 Gy-fraction required 3.9 min (RapidArc), 11.6 min (conf-SBRT), and 12 min (IMRT)., Conclusions: In stage I lung tumors measuring <70 cm(3), RapidArc plans achieved both the highest dose conformity and shortest delivery times., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Urinary tract infections (UTIs) are among the most common infectious diseases of humans, with Escherichia coli being responsible for >80% of all cases. Asymptomatic bacteriuria (ABU) occurs when bacteria colonize the urinary tract without causing clinical symptoms and can affect both catheterized patients (catheter-associated ABU [CA-ABU]) and noncatheterized patients. Here, we compared the virulence properties of a collection of ABU and CA-ABU nosocomial E. coli isolates in terms of antibiotic resistance, phylogenetic grouping, specific UTI-associated virulence genes, hemagglutination characteristics, and biofilm formation. CA-ABU isolates were similar to ABU isolates with regard to the majority of these characteristics; exceptions were that CA-ABU isolates had a higher prevalence of the polysaccharide capsule marker genes kpsMT II and kpsMT K1, while more ABU strains were capable of mannose-resistant hemagglutination. To examine biofilm growth in detail, we performed a global gene expression analysis with two CA-ABU strains that formed a strong biofilm and that possessed a limited adhesin repertoire. The gene expression profile of the CA-ABU strains during biofilm growth showed considerable overlap with that previously described for the prototype ABU E. coli strain, 83972. This is the first global gene expression analysis of E. coli CA-ABU strains. Overall, our data suggest that nosocomial ABU and CA-ABU E. coli isolates possess similar virulence profiles.