A targeted amplicon sequencing panel to simultaneously identify mosquito species and Plasmodium presence across the entire Anopheles genus.

Anopheles is a diverse genus of mosquitoes comprising over 500 described species, including all known human malaria vectors. While a limited number of key vector species have been studied in detail, the goal of malaria elimination calls for surveillance of all potential vector species. Here, we develop a multilocus amplicon sequencing approach that targets 62 highly variable loci in the Anopheles genome and two conserved loci in the Plasmodium mitochondrion, simultaneously revealing both the mosquito species and whether that mosquito carries malaria parasites. We also develop a cheap, nondestructive, and high-throughput DNA extraction workflow that provides template DNA from single mosquitoes for the multiplex PCR, which means specimens producing unexpected results can be returned to for morphological examination. Over 1000 individual mosquitoes can be sequenced in a single MiSeq run, and we demonstrate the panel's power to assign species identity using sequencing data for 40 species from Africa, Southeast Asia, and South America. We also show that the approach can be used to resolve geographic population structure within An. gambiae and An. coluzzii populations, as the population structure determined based on these 62 loci from over 1000 mosquitoes closely mirrors that revealed through whole genome sequencing. The end-to-end approach is quick, inexpensive, robust, and accurate, which makes it a promising technique for very large-scale mosquito genetic surveillance and vector control.

Genetic surveillance in the Greater Mekong subregion and South Asia to support malaria control and elimination.

Background: National Malaria Control Programmes (NMCPs) currently make limited use of parasite genetic data. We have developed GenRe-Mekong, a platform for genetic surveillance of malaria in the Greater Mekong Subregion (GMS) that enables NMCPs to implement large-scale surveillance projects by integrating simple sample collection procedures in routine public health procedures. Methods: Samples from symptomatic patients are processed by SpotMalaria, a high-throughput system that produces a comprehensive set of genotypes comprising several drug resistance markers, species markers and a genomic barcode. GenRe-Mekong delivers Genetic Report Cards, a compendium of genotypes and phenotype predictions used to map prevalence of resistance to multiple drugs. Results: GenRe-Mekong has worked with NMCPs and research projects in eight countries, processing 9623 samples from clinical cases. Monitoring resistance markers has been valuable for tracking the rapid spread of parasites resistant to the dihydroartemisinin-piperaquine combination therapy. In Vietnam and Laos, GenRe-Mekong data have provided novel knowledge about the spread of these resistant strains into previously unaffected provinces, informing decision-making by NMCPs. Conclusions: GenRe-Mekong provides detailed knowledge about drug resistance at a local level, and facilitates data sharing at a regional level, enabling cross-border resistance monitoring and providing the public health community with valuable insights. The project provides a rich open data resource to benefit the entire malaria community. Funding: The GenRe-Mekong project is funded by the Bill and Melinda Gates Foundation (OPP11188166, OPP1204268). Genotyping and sequencing were funded by the Wellcome Trust (098051, 206194, 203141, 090770, 204911, 106698/B/14/Z) and Medical Research Council (G0600718). A proportion of samples were collected with the support of the UK Department for International Development (201900, M006212), and Intramural Research Program of the National Institute of Allergy and Infectious Diseases.

Altered Doppler flow patterns in cirrhosis patients: an overview.

Doppler ultrasonography of the hepatic vasculature is an integral part of evaluating precirrhotic and cirrhotic patients. While the reversal of the portal venous flow is a well-recognized phenomenon, other flow patterns, although not as easily understood, may play an important role in assessing the disease status. This article discusses the different characteristic flow patterns observed from the portal vein, hepatic artery, and hepatic vein in patients with liver cirrhosis or related complications and procedures. Knowledge of these different flow patterns provides additional information that may reinforce the diagnosis of cirrhosis, help in staging, and offer prognostic information for determining the direction of therapy. Doppler ultrasonography is invaluable when liver transplantation is being considered and aids in the diagnosis of cirrhosis and portal hypertension.

Duplex Doppler Imaging of Dialysis Fistulae and Grafts.

Arteriovenous fistulae and grafts for hemodialysis access are a lifeline in patients with end-stage renal disease. A significant cause of morbidity and mortality in this population is dialysis access dysfunction. Duplex ultrasound imaging is an excellent modality to evaluate arteriovenous fistulae and grafts, the 2 main types of long-term hemodialysis access. This review provides a detailed Doppler ultrasound protocol for evaluation of fistulae or grafts to familiarize imagers with their normal appearance, highlighting common dialysis access complications.

Chemotherapy-related complications in the kidneys and collecting system: an imaging perspective.

UNLABELLED: Nephrotoxicity is a common adverse effect of many chemotherapeutic agents. The agents most commonly associated with chemotherapy-associated nephrotoxicity are methotrexate, semustine, streptozocin, mithramycin, and cisplatin. Certain chemotherapeutic agents have adverse effects on the kidneys and urothelium that can be visualized radiographically, including cystic change, interstitial nephritis, papillary necrosis, urothelial changes, haemorrhagic cystitis, acute tubular necrosis, and infarction. This review focuses on imaging features identifying complications of chemotherapy in the kidneys and collecting system and provides didactic cases to alert referring clinicians. TEACHING POINTS: • Nephrotoxicity is a common adverse effect of many chemotherapeutic agents. • Chemotherapies have adverse renal and urothelial effects that can be visualized radiographically. • Crizotinib use can result in the development of complex renal cysts.

Interventional radiology delivers high-value health care and is an Imaging 3.0 vanguard.

Given the changing climate of health care and the imperative to add value, radiologists must join forces with the rest of medicine to deliver better patient care in a more cost-effective, evidence-based manner. For several decades, interventional radiology has added value to the health care system through innovation and the provision of alternative and effective minimally invasive treatments, which have decreased morbidity, mortality, and overall cost. The clinical practice of interventional radiology embodies many of the features of Imaging 3.0, the program recently launched by the ACR. We provide a review of some of the major contributions made by interventional radiology and offer general principles from that experience, which are applicable to all radiologists.

Dedifferentiation of cancer cells following recovery from a potentially lethal damage is mediated by H2S-Nampt.

Recently, we reported that cancer cells that recover from a potentially lethal damage gain new phenotypic features comprised of mitochondrial structural remodeling associated with increased glycolytic dependency and drug resistance. Here, we demonstrate that a subset of cancer cells, upon recovery from a potentially lethal damage, undergo dedifferentiation and express genes, which are characteristic of undifferentiated stem cells. While these cells are competent in maintaining differentiated progeny of tumor, they also exhibit transdifferentiation potential. Dedifferentiation is characterized by accumulation of hydrogen sulfide (H2S), which triggers up-regulation of nicotinamide phosphoribosyltransferase (Nampt) accompanied by changes in the redox state. The molecular events triggered by Nampt include elevated production of NAD(+) and up-regulation of H2S producing enzymes, cystathionine beta synthase (CBS) and cystathionase (CTH) with 3-mercaptopyruvate sulfurtransferase (MST) being detectable only in 3D spheroids. Suppression of Nampt, or inactivation of H2S producing enzymes, all reduce H2S production and reverse the ability of cells to dedifferentiate. Moreover, H2S induced stem cell markers in parental cancer cells in a manner similar to that observed in damage recovered cells. These data suggest of existence of a positive feedback loop between H2S and Nampt that controls dedifferentiation in cancer cells that recover from a potentially lethal damage.

A H2S-Nampt dependent energetic circuit is critical to survival and cytoprotection from damage in cancer cells.

We recently demonstrated that cancer cells that recover from damage exhibit increased aerobic glycolysis, however, the molecular mechanism by which cancer cells survive the damage and show increased aerobic glycolysis remains unknown. Here, we demonstrate that diverse cancer cells that survive hypoxic or oxidative damage show rapid cell proliferation, and develop tolerance to damage associated with increased production of hydrogen sulfide (H2S) which drives up-regulation of nicotinamide phosphoribosyltransferase (Nampt). Consistent with existence of a H2S-Nampt energetic circuit, in damage recovered cancer cells, H2S, Nampt and ATP production exhibit a significant correlation. Moreover, the treatment of cancer cells with H2S donor, NaHS, coordinately increases Nampt and ATP levels, and protects cells from drug induced damage. Inhibition of cystathionine beta synthase (CBS) or cystathionase (CTH), enzymes which drive generation of H2S, decreases Nampt production while suppression of Nampt pathway by FK866, decreases H2S and ATP levels. Damage recovered cells isolated from tumors grown subcutaneously in athymic mice also show increased production of H2S, Nampt and ATP levels, associated with increased glycolysis and rapid proliferation. Together, these data show that upon recovery from potential lethal damage, H2S-Nampt directs energy expenditure and aerobic glycolysis in cancer cells, leads to their exponential growth, and causes a high degree of tolerance to damage. Identification of H2S-Nampt as a pathway responsible for induction of damage tolerance in cancer cells may underlie resistance to therapy and offers the opportunity to target this pathway as a means in treatment of cancer.

Computer-aided diagnosis of hyperacute stroke with thrombolysis decision support using a contralateral comparative method of CT image analysis.

New and improved techniques have been continuously introduced into CT and MR imaging modalities for the diagnosis and therapy planning of acute stroke. Nevertheless, non-contrast CT (NCCT) is almost always used by every institution as the front line diagnostic imaging modality due to its high affordability and availability. Consequently, the potential reward of extracting as much clinical information as possible from NCCT images can be very great. Intravenous tissue plasminogen activator (tPA) has become the gold standard for treating acute ischemic stroke because it is the only acute stroke intervention approved by the FDA. ASPECTS scoring based on NCCT images has been shown to be a reliable scoring method that helps physicians to make sound decisions regarding tPA administration. In order to further reduce inter-observer variation, we have developed the first end-to-end automatic ASPECTS scoring system using a novel method of contralateral comparison. Due to the self-adaptive nature of the method, our system is robust and has good generalizability. ROC analysis based on evaluation of 103 subjects who presented to the stroke center of Chang Gung Memorial Hospital with symptoms of acute stroke has shown that our system's dichromatic classification of patients into thrombolysis indicated or thrombolysis contraindicated groups has achieved a high accuracy rate with AUC equal to 90.2 %. The average processing time for a single case is 170 s. In conclusion, our system has the potential of enhancing quality of care and providing clinical support in the setting of a busy stroke or emergency center.

Centromere tethering confines chromosome domains.

The organization of chromosomes into territories plays an important role in a wide range of cellular processes, including gene expression, transcription, and DNA repair. Current understanding has largely excluded the spatiotemporal dynamic fluctuations of the chromatin polymer. We combine in vivo chromatin motion analysis with mathematical modeling to elucidate the physical properties that underlie the formation and fluctuations of territories. Chromosome motion varies in predicted ways along the length of the chromosome, dependent on tethering at the centromere. Detachment of a tether upon inactivation of the centromere results in increased spatial mobility. A confined bead-spring chain tethered at both ends provides a mechanism to generate observed variations in local mobility as a function of distance from the tether. These predictions are realized in experimentally determined higher effective spring constants closer to the centromere. The dynamic fluctuations and territorial organization of chromosomes are, in part, dictated by tethering at the centromere.

Dipeptidyl peptidase IV inhibition does not adversely affect immune or virological status in HIV infected men and women: a pilot safety study.

CONTEXT: People infected with HIV have a higher risk for developing insulin resistance, diabetes, and cardiovascular disease than the general population. Dipeptidyl peptidase IV (DPP4) inhibitors are glucose-lowering medications with pleiotropic actions that may particularly benefit people with HIV, but the immune and virological safety of DPP4 inhibition in HIV is unknown. OBJECTIVE: DPP4 inhibition will not reduce CD4+ T lymphocyte number or increase HIV viremia in HIV-positive adults. DESIGN: This was a randomized, placebo-controlled, double-blind safety trial of sitagliptin in HIV-positive adults. SETTING: The study was conducted at an academic medical center. PARTICIPANTS: Twenty nondiabetic HIV-positive men and women (9.8 ± 5.5 years of known HIV) taking antiretroviral therapy and with stable immune (625 ± 134 CD4+ T cells per microliter) and virological (<48 copies HIV RNA per milliliter) status. INTERVENTION: The intervention included sitagliptin (100 mg/d) vs matching placebo for up to 24 weeks. MAIN OUTCOME MEASURES: CD4+ T cell number and plasma HIV RNA were measured every 4 weeks; fasting serum regulated upon activation normal T-cell expressed and secreted (RANTES), stromal derived factor (SDF)-1α, Soluble TNF receptor II, and oral glucose tolerance were measured at baseline, week 8, and the end of study. ANOVA was used for between-group comparisons; P < .05 was considered significant. RESULTS: Compared with placebo, sitagliptin did not reduce CD4+ T cell count, plasma HIV RNA remained less than 48 copies/mL, RANTES and soluble TNF receptor II concentrations did not increase. SDF1α concentrations declined (P < .0002) in the sitagliptin group. The oral glucose tolerance levels improved in the sitagliptin group at week 8. CONCLUSIONS: Despite lowering SDF1α levels, sitagliptin did not adversely affect immune or virological status, or increase immune activation, but did improve glycemia in healthy, nondiabetic HIV-positive adults. These safety data allow future efficacy studies of sitagliptin in HIV-positive people with cardiometabolic complications.

A Rare but Significant Cause of an Enlarging Neck Mass: A Case Report of Primary Thyroid Lymphoma.

Primary thyroid lymphoma is a rare form of non-Hodgkin's lymphoma that typically presents as a rapidly enlarging goiter in patients with a history of autoimmune thyroid disease. We present the evaluation, pitfalls in diagnosis, and treatment of a 49-year-old woman with diffuse large B-cell primary thyroid lymphoma presenting with airway obstruction.

Comparison of Sexual Dysfunction Using the Female Sexual Function Index following Surgical Treatments for Uterine Fibroids.

Uterine fibroids are a common problem in women. Statistics showing 20-50% of fibroids produce symptoms and consequently patients seek surgical intervention to improve their quality of life. Treatments for fibroids are typically successful in controlling the fibroid disease, yet sexual function following invasive surgical treatments for fibroids can be jeopardized. The Sexual Function Index (FSFI) is a valid instrument producing quantifiable reproducible results. In this paper three case reports are evaluated by the FSFI and compared between the following treatment groups: hysterectomy, myomectomy, and uterine embolization. Our goal is to illustrate how each of these treatment outcomes can result in sexual dysfunction and therefore decreased quality of life. Effects of invasive fibroid treatments on sexual functioning would be helpful in guiding patient's ultimate decisions regarding treatment.

Basal ganglia structures differentially contribute to verbal fluency: evidence from Human Immunodeficiency Virus (HIV)-infected adults.

BACKGROUND: The basal ganglia (BG) are involved in executive language functions (i.e., verbal fluency) through their connections with cortical structures. The caudate and putamen receive separate inputs from prefrontal and premotor cortices, and may differentially contribute to verbal fluency performance. We examined BG integrity in relation to lexico-semantic verbal fluency performance among older HIV infected adults. METHOD: 20 older (50+ years) HIV+ adults underwent MRI and were administered measures of semantic and phonemic fluency. BG (caudate, putamen) regions of interest were extracted. RESULTS: Performance on phonemic word generation significantly predicted caudate volume, whereas performance on phonemic switching predicted putamen volume. CONCLUSIONS: These findings suggest a double dissociation of BG involvement in verbal fluency tasks with the caudate subserving word generation and the putamen associated with switching. As such, verbal fluency tasks appear to be selective to BG function.

Lower-extremity endovascular interventions for Medicare beneficiaries: comparative effectiveness as a function of provider specialty.

PURPOSE: Lower-extremity endovascular interventions are increasingly being performed by vascular surgeons (VSs) and interventional cardiologists (ICs) in addition to interventional radiologists (IRs). Regardless of specialty, well trained, experienced, and dedicated operators are expected to offer the best outcomes. To examine specialty-specific trends, outcomes of percutaneous lower-extremity revascularizations in Medicare beneficiaries were compared according to physician specialty types providing the service. MATERIALS AND METHODS: Medicare Standard Analytical Files that contain longitudinal data of all services (physician, inpatient, outpatient) provided to a 5% sample of Medicare beneficiaries were studied. All claims for percutaneous angioplasty, atherectomy, and stent implantation of lower-extremity arteries during the years 2005­2007 were extracted, and the following outcomes were assessed: mortality, transfusion, intensive care unit (ICU) use, length of stay, and subsequent revascularization or amputation. Outcomes were compared by using regression models adjusted for age, sex, race, emergency department admission, and comorbid conditions. RESULTS: Most outcomes were significantly worse if the service was provided by vascular surgeons compared with other vascular specialists. The in-hospital mortality rate for procedures performed by VSs was 19% higher than for those performed by others, but this difference was not significant (P =.351). Adjusted average 1-year procedure costs were significantly lower for IRs ($17,640) than for VSs ($19,012) or ICs ($19,096). CONCLUSIONS: Medicare data show that endovascular lower-extremity revascularization by vascular surgeons results in more transfusion and ICU use, longer hospital stay, more repeat revascularization procedures or amputations, and higher costs compared with procedures performed by interventional radiologists.

Associated vascular injury in patients with bile duct injury during cholecystectomy.

Cholecystectomy remains one of the most commonly performed procedures in general surgery. Although the incidence, diagnosis, and treatment of bile duct (BD) injuries have been well described, studies characterizing associated vascular injuries are limited. The objective of this study was to analyze the frequency and management of associated vascular and BD injury after cholecystectomy. A total of 50 patients were referred to a tertiary institution for BD injuries from 1996 to 2010. Thirty-nine (78%) of the patients were female with the mean age of 49 years (range, 14 to 86 years). Seventy-five per cent of the injuries were Strasberg Type E. Nine patients (18%) had associated vascular injuries. Six patients had injuries to the right hepatic artery; in one patient, both the right and left hepatic arteries were damaged. Five patients had right portal vein injuries; three of these subsequently died. In conclusion, as a result of the high incidence of associated vascular injury, a thin-collimation CT angiogram and/or mesenteric angiogram with portal venous imaging should be considered as part of the preoperative evaluation in patients with BD injury.