Short-Term Dynamics of Bdellovibrio and Like Organisms in Lake Geneva in Response to a Simulated Climatic Extreme Event.

The short time-scale dynamics of three families of Bdellovibrio and like organisms (i.e. Bdellovibrionaceae, Peredibacteraceae, and Bacteriovoracaceae) were studied on the surface waters of Lake Geneva in summer. Using mesocosms deployed nearshore in July 2019, we simulated an extreme climatic event (an input of carbon from the watershed in response to runoff from the catchment, light reduction, and mixing in response to stormy conditions) and aimed to study the impact of both abiotic and biotic factors on their dynamics. The three families of Bdellovibrio and like organisms (BALOs) showed different dynamics during the experiment. Peredibacteraceae was the most abundant group, whereas Bacteriovoracaceae was the least abundant. Compared with the other two families, the abundance of Bdellovibrionaceae did not fluctuate, remaining relatively stable over time. Environmental variables only partially explained the dynamics of these families; in particular, temperature, pH, and chloride concentrations were positively correlated with Bacteriovoracaceae, Bdellovibrionaceae, and Peredibacteraceae abundance, respectively. Prokaryote-like particles (PLPs), such as those with high DNA content (HDNA), were strongly and positively correlated with Peredibacteraceae and Bacteriovoracaceae. In contrast, no relationships were found between Bdellovibrionaceae and PLP abundance, nor between the virus-like particles (VLPs) and the different BALOs. Overall, the experiment revealed that predation was stable in the face of the simulated climatic events. In addition, we observed that Peredibacteraceae and Bacteriovoracaceae share common traits, while Bdellovibrionaceae seems to constitute a distinct category.

Low effective population sizes in Amblyomma variegatum, the tropical bont tick.

Effective population sizes have rarely been estimated in ticks despite the importance of this parameter for evaluating the evolutionary and adaptive potential of tick populations. The present study was aimed at evaluating the effective population sizes of Amblyomma variegatum, the tropical bont tick, in three villages in Burkina Faso. For this purpose, microsatellites markers were developed. Eight out of 19 assessed markers provided good amplification results with 4 to 24 alleles recorded per marker on 216 genotyped ticks. The within-samples polymorphism was congruent with Hardy-Weinberg expectations at four markers while sex linkage and/or null alleles were observed at the others. As sampling involved two tick generations, effective population sizes were independently estimated by two methods insensitive to heterozygosity: the first one is based on linkage disequilibrium analysis within a single cohort while the second uses the changes in allele frequencies across generations. Both methods estimated the number of reproducing ticks ranging from two to a few tens reproductive adults per village and cohort. Such small estimates are congruent with the rarity of records of acaricide resistance in A. variegatum.

Outcomes following a negative computed tomography pulmonary angiography according to pulmonary embolism prevalence: a meta-analysis of the management outcome studies.

Essentials Computed tomographic pulmonary angiography (CTPA) is used to exclude pulmonary embolism. This meta-analysis explores the occurrence of venous thromboembolic events (VTE) after a CTPA. Occurrence of VTE after a negative CTPA is ˜8% in study subgroups with a prevalence of PE ≥ 40%. CTPA may be insufficient to safely rule out VTE as a stand-alone diagnostic test for this subgroup. SUMMARY: Background Outcome studies have reported the safety of computed tomographic pulmonary angiography (CTPA) as a stand-alone imaging technique to rule out pulmonary embolism (PE). Whether this can be applied to all clinical probabilities remains controversial. Objectives We performed a meta-analysis to determine the proportion of patients with venous thromboembolic events (VTE) despite a negative CTPA according to pretest PE prevalence. Methods We searched MEDLINE, EMBASE and the Cochrane Library (January 1990 to May 2017) for outcome studies recruiting patients with suspected PE using CTPA as a diagnostic strategy. The primary outcome was the cumulative occurrence of VTE at 3 months following a negative CTPA. Results Twenty-two different studies were identified. VTE was confirmed in 2.4% of patients (95% CI, 1.3-3.8%) either at the time of the index event or in the 3 months follow-up. Subgroup analyses suggested that the cumulative occurrence of VTE was related to pretest prevalence of PE, as VTE occurred in 1.8% (95% CI, 0.5-3.7%), 1.4% (95% CI, 0.7-2.3%), 1.0% (95% CI, 0.5-1.8%) and 8.1% (95% CI, 3.5-14.5%) of subgroups of patients with a PE prevalence < 20%, 20-29%, 30-39% and ≥ 40%, respectively. This was further confirmed using meta-regression analysis. Conclusions The negative predictive value of CTPA for VTE varies according to pretest prevalence of PE, and is likely to be insufficient to safely rule out VTE as a stand-alone diagnostic test amongst patients at the highest pretest probability of VTE. Prospective studies are required to validate the appropriate diagnostic algorithm for this subgroup of patients.

[Oncologic and perioperative outcomes of robot-assisted radical cystectomy: A single-center prospective series]. / Résultats oncologiques et périopératoires de la cystectomie totale robot-assistée pour cancer : une série prospective monocentrique.

INTRODUCTION: The aim of this study was to report perioperative and oncological outcomes of robot-assisted radical cystectomy (RARC) in a single-center series and to evaluate the impact of the experience on perioperative outcomes. METHODS: Between March 2012 and January 2016, 41 patients underwent RARC associated with extended pelvic lymphadenectomy for muscle-invasive bladder cancer. All RARC included were performed by a single-surgeon in one center. Perioperative and oncological datas were collected prospectively. Recurrence-free (RFS), overall (OS) and cancer-specific survivals (CSS) were estimated using the Kaplan-Meier Method. The impact of the experience on perioperative data was estimated using Spearman's correlation test. RESULTS: Mean age was 67,7years (±10.6). Most patients underwent neoadjuvant chemotherapy (73.2%). Mean operative time and mean estimated blood loss were respectively 319.5minutes (±85.3) and 662.5mL (±360.9). Eight patients needed perioperative blood transfusion (19.5%). Conversion to open surgery was necessary in 3 cases (7.3%). Ileal neobladder was performed in 26.8% of the cases (54.5% being performed intracorporeal), and non-continent urinary diversion in 73.2%. Mean nodal yield was 17.7 (±9.3). Positive surgical margins were observed in 1 patient (2.3%). Mean length of stay was 13.2 days (±9.8). Postoperative complication rate was 46,3%. After a median follow-up of 16months, estimated 2 year-OS and CSS were respectively 62 and 76.1%. Estimated 2-year RFS was 67.6%. Perioperative outcomes improved with experience with a significant decrease in operative time (P=0.04) and a significant increase of nodal yield (P=0.05). CONCLUSION: In this single-center prospective study, satisfactory perioperative and oncological outcomes after RARC were observed despite the learning curve. Perioperative outcomes improved with surgeon's experience. Further studies are needed to confirm these findings. LEVEL OF EVIDENCE: 4.

[Pathological advances in renal, prostatic, bladder and testis neoplasia]. / Actualités en pathologie tumorale rénale, prostatique, vésicale et testiculaire.

INTRODUCTION: The ISUP (International Society of Urological Pathology) Consensus Conferences between 2012 and 2015 made recommendations regarding the classification, staging, prognostic factors of adult tumors from kidney, prostate, bladder and testis. The main points of these recommendations are highlighted in this article. MATERIALS AND METHODS: This article is based on a systematic literature search by using different keywords "cancer, kidney, prostate, bladder, testis, pathology, classification" from Pubmed database. Only publications between 2012 and 2015 were retained. RESULTS: The different Consensus conferences since 2012 in uropathology have provided international guidelines for the classification, grading and staging of tumors in kidney, bladder, prostate and testis. We identified in this article the main points of these new guidelines that are about to be published in the new 2016 WHO classification of urogenital tract tumors in adult. CONCLUSION: New pathological guidelines in urogenital tumors have to be taken into account for a better diagnosis and therapy.

[Impact of robotic assistance on the use and the outcomes of nephron-sparing surgery: A single center experience]. / Impact de l'assistance robotique sur la pratique et les résultats de la chirurgie rénale conservatrice : une expérience monocentrique.

INTRODUCTION: The aim of this study was to assess the impact of the acquisition of a Da Vinci® robot on the use and outcomes of partial nephrectomy (PN). PATIENTS AND METHODS: It was a single center retrospective study including 280 patients who underwent PN from January 2006 to May 2013. The number of PN, tumors and patients' characteristics and perioperative outcomes have been assessed over 3 periods defined according to the main surgical approach: 2006-2008 (open PN), 2008-2010 (laparoscopic PN) and 2010-2013 (robotic PN). RESULTS: Over the study period, the surgical approach has changed significantly in favor of minimally-invasive surgery and especially robotic approach. The PN/nephrectomy rate has also evolved to a higher proportion of PN over radical nephrectomy (P=0.002). No significant difference was noted between the three periods in terms of tumor size but there was a higher rate of highly complex tumors (RENAL score≥10) during the last period (10.7%; 18.6% and 33.2%; P=0.04). Warm ischemia time increased from 2006-2008 to 2008-2010 (26 vs. 23minutes) but decreased thereafter during the robotic era (14.5minutes; P<0.001). Regarding postoperative outcomes, the only change was a decreased length of stay over time (P=0.003). CONCLUSION: In this single center series, the robotic approach was associated with a spread of PN and with an improvement of nephron-sparing surgery outcomes. LEVEL OF EVIDENCE: 4.

First Evidence of an Important Organic Matter Trophic Pathway between Temperate Corals and Pelagic Microbial Communities.

Mucus, i.e., particulate and dissolved organic matter (POM, DOM) released by corals, acts as an important energy carrier in tropical ecosystems, but little is known on its ecological role in temperate environments. This study assessed POM and DOM production by the temperate coral Cladocora caespitosa under different environmental conditions. The subsequent enzymatic degradation, growth of prokaryotes and virus-like particles (VLPs) as well as changes in the structure of the prokaryotic communities were also monitored. C. caespitosa produced an important quantity of mucus, which varied according to the environmental conditions (from 37.8 to 67.75 nmol carbon h-1 cm-2), but remained higher or comparable to productions observed in tropical corals. It has an important nutritional value, as highlighted by the high content in dissolved nitrogen (50% to 90% of the organic matter released). Organic matter was rapidly degraded by prokaryotes' enzymatic activities, and due to its nitrogen content, aminopeptidase activity was 500 fold higher than the α-glucosidase activity. Prokaryotes, as well as VLPs, presented a rapid growth in the mucus, with prokaryote production rates as high as 0.31 µg h-1 L-1. Changes in bacterial and archaeal communities were observed in the ageing mucus and between mucus and the water column, suggesting a clear impact of mucus on microorganism diversity. Overall, our results show that the organic matter released by temperate corals, such as C. caespitosa, which can form reef structures in the Mediterranean Sea, stimulates microbial activity and thereby functions as a significant carbon and nitrogen supplier to the microbial loop.

Colonization of the Mediterranean basin by the vector biting midge species Culicoides imicola: an old story.

Understanding the demographic history and genetic make-up of colonizing species is critical for inferring population sources and colonization routes. This is of main interest for designing accurate control measures in areas newly colonized by vector species of economically important pathogens. The biting midge Culicoides imicola is a major vector of orbiviruses to livestock. Historically, the distribution of this species was limited to the Afrotropical region. Entomological surveys first revealed the presence of C. imicola in the south of the Mediterranean basin by the 1970s. Following recurrent reports of massive bluetongue outbreaks since the 1990s, the presence of the species was confirmed in northern areas. In this study, we addressed the chronology and processes of C. imicola colonization in the Mediterranean basin. We characterized the genetic structure of its populations across Mediterranean and African regions using both mitochondrial and nuclear markers, and combined phylogeographical analyses with population genetics and approximate Bayesian computation. We found a west/east genetic differentiation between populations, occurring both within Africa and within the Mediterranean basin. We demonstrated that three of these groups had experienced demographic expansions in the Pleistocene, probably because of climate changes during this period. Finally, we showed that C. imicola could have colonized the Mediterranean basin in the Late Pleistocene or Early Holocene through a single event of introduction; however, we cannot exclude the hypothesis involving two routes of colonization. Thus, the recent bluetongue outbreaks are not linked to C. imicola colonization event, but rather to biological changes in the vector or the virus.

Assessment of the safety and immunogenicity of Rhodococcus equi-secreted proteins combined with either a liquid nanoparticle (IMS 3012) or a polymeric (PET GEL A) water-based adjuvant in adult horses and foals--identification of promising new candidate antigens.

Rhodococcus equi is the most common infectious cause of mortality in foals between 1 and 6 months of age. Because of an increase in the number of antibiotic-resistant strains, the optimization of a prophylactic strategy is a key factor in the comprehensive management of R. equi pneumonia. The objectives of this study were to assess the safety and immunogenicity of R. equi-secreted proteins (ReSP) co-administered with either the nanoparticular adjuvant Montanide™ IMS 3012 VG, or a new polymeric adjuvant Montanide™ PET GEL A, and to further investigate the most immunogenic proteins for subsequent immunization/challenge experiments in the development of a vaccine against rhodoccocal pneumonia. The approach involved two phases. The first phase aimed to investigate the safety of vaccination in six adult horses. The second phase aimed to determine the safety and immunogenicity of vaccination in twelve 3-week-old foals. We set out to develop a method based on ultrasound measurements for safety assessment in adult horses in order to evaluate any in situ changes at the injection site, in the skin or the underlying muscle, with quantitative and qualitative data revealing that administration of ReSP combined with the Pet Gel A adjuvant led to an increase in local inflammation, associated with 4- to 7-fold higher levels of anti-R. equi IgGa, IgGb and IgGT, compared to administration of ReSP associated with IMS 3012 adjuvant, but without any impact on animal demeanor. Investigations were then performed in foals with serological and clinical follow-up until 6 months of age. Interestingly, we observed in foals a much lower incidence of adverse local tissue reactions at the injection site than in adult horses, with transient and moderate swelling for the group that received ReSP combined with Pet Gel A. Immunized foals with Pet Gel A adjuvant exhibited a similar response in both IgGa and IgGT levels, but a lower response in IgGb levels, compared to adult horses, with a subisotype profile that may however reflect a bias favorable to R. equi resistance. From the crude extract of secreted proteins, dot-blot screening enabled identification of cholesterol oxidase, mycolyl transferase 3, and PSP (probable secreted protein) as the most immunogenic candidates. Taken together, these results are encouraging in developing a vaccine for foals.

Evolution of radiological findings detected in the limbs of 321 young horses between the ages of 6 and 18 months.

The objective of this study was to evaluate the spontaneous evolution of radiological findings (RF) indicative of juvenile osteochondral conditions (JOCC) in a field study. A radiographic survey was performed at the age of 6 and 18months in 321 young horses from three breeds (French Trotter Standardbred, Selle-Français Warmblood and Thoroughbred). Each RF identified at 6months was re-evaluated at 18months, and classified as (1) disappeared, (2) improved, (3) stable, (4) deteriorated or (5) new when it was not identified at 6months. Only 32.3% of all RF identified at the age of 6months were stable, while at 18months 46.6% had disappeared and 38.7% were new radiological abnormalities. Evolution of RF varied according to the joint involved and the type of lesion. In the stifle, 85.7% of the RF of osteochondral fragmentation (OCF) of the lateral ridge of the femoral trochlea regressed (disappeared or improved). In the hock, 53.1% of the RF of OCD of the intermediate ridge of the tibial cochlea were stable. In the fetlock, dorsal and plantar OCF were frequently observed as new radiological abnormalities at 18months (51% of the dorsal RF and 45.8% of the plantar). As many lesions develop or change after 6months, a definitive radiographic assessment of the JOCC status is more reliable at 18months. Knowing the spontaneous healing capacity of some lesions is useful in helping to decide the appropriate management.

Radiographic findings of juvenile osteochondral conditions detected in 392 foals using a field radiographic protocol.

Radiographic examination in young horses is commonly used to detect juvenile osteochondral conditions (JOCC). The aim of this study is to present the radiographic findings (RF) detected at the age of 6months in the joints of 392 foals from three breeds: French Trotter Standardbreds (FT), Selle-Français Warmbloods (SF) and Thoroughbreds (TB). Radiographic examination included lateromedial views of the fore and hind digits, tarsi and stifles, and dorsopalmar projections of both carpi. In all areas RF indicative of JOCC were classified according to a four-grade severity scoring system. Distribution and severity of the RF varied according to breed with the SF most affected. The most frequently affected site was the hind fetlock (28.3% of foals). The second most affected site was the fore fetlock in TB and SF, while the carpus and proximal tarsus were the second and third affected sites in FT. The radiographic protocol and grading system used can serve as a basis for field research and further studies on the evolution of RF, risk factors for JOCC or the correlation between RF and the subsequent athletic career of horses.

Phage adsorption to bacteria in the light of the electrostatics: a case study using E. coli, T2 and flow cytometry.

The addition of sodium chloride to freshwater or diluted minimal salt medium increases the adsorption of T2 phages on Escherichia coli. For the first time the adsorption in diluted minimal salt medium was measured by counting unadsorbed phages (i.e. free particles) using flow cytometry, allowing a gentle separation between adsorbed and unadsorbed phages. Flow cytometry was able to detect weakly adsorbed phage that remained undetected using classical centrifugation-based methods and this allowed us to show that increasing ionic strength enhances the phage adsorption to its bacterial host with an extremely low detection limit. A key result was that the adsorption in high ionic strength (i.e. 100 mM) reached 4.5±0.1×10⁻5 mL/min which is 1400 fold higher than previously reported values. In order to understand the mechanism underpinning such a weak phage adsorption, the zeta potentials and the diffusion coefficient of the particles were measured by dynamic light scattering. The bacterial cells and the phages had zeta potentials between -60 mV and -10 mV and -30 mV and -10 mV, respectively. The diffusion coefficient of the phage was 2.8±0.4×10⁻¹² m² s⁻¹ corresponding to a hydrodynamic radius of 104±15 nm. However significant adsorption occurs in conditions where the DLVO theory predicts that minimal encounter, suggesting that forces other that electrostatic repulsion and Van der Waals interaction (e.g. potential impurities, particle shape and other biological characteristics) are likely to interplay.

Beyond an AFLP genome scan towards the identification of immune genes involved in plague resistance in Rattus rattus from Madagascar.

Genome scans using amplified fragment length polymorphism (AFLP) markers became popular in nonmodel species within the last 10 years, but few studies have tried to characterize the anonymous outliers identified. This study follows on from an AFLP genome scan in the black rat (Rattus rattus), the reservoir of plague (Yersinia pestis infection) in Madagascar. We successfully sequenced 17 of the 22 markers previously shown to be potentially affected by plague-mediated selection and associated with a plague resistance phenotype. Searching these sequences in the genome of the closely related species Rattus norvegicus assigned them to 14 genomic regions, revealing a random distribution of outliers in the genome (no clustering). We compared these results with those of an in silico AFLP study of the R. norvegicus genome, which showed that outlier sequences could not have been inferred by this method in R. rattus (only four of the 15 sequences were predicted). However, in silico analysis allowed the prediction of AFLP markers distribution and the estimation of homoplasy rates, confirming its potential utility for designing AFLP studies in nonmodel species. The 14 genomic regions surrounding AFLP outliers (less than 300 kb from the marker) contained 75 genes encoding proteins of known function, including nine involved in immune function and pathogen defence. We identified the two interleukin 1 genes (Il1a and Il1b) that share homology with an antigen of Y. pestis, as the best candidates for genes subject to plague-mediated natural selection. At least six other genes known to be involved in proinflammatory pathways may also be affected by plague-mediated selection.

[Tropical sprue in an expatriate]. / Sprue tropicale chez un expatrié.

INTRODUCTION: Tropical sprue is a postinfective malabsorption syndrome that occurs in some tropical endemic areas. CASE REPORT: A 65-year-old Caucasian patient, with no significant past medical history, living in Cambodia for 10 years, presented with a 23 kg weight loss and chronic diarrhea. Clinical examination was unremarkable. Laboratory tests showed a moderate nutritional deficiency syndrome. The upper gastrointestinal endoscopy showed duodenal villous atrophy and histological analysis confirmed subtotal villous atrophy with important intraepithelial lymphocytosis. The diagnosis of tropical sprue was considered on the epidemiological, clinical and biological context, and the absence of other cause of villous atrophy. A three-month duration treatment with antibiotics, folic acid and vitamin B12 was initiated. The clinical course was favorable with disappearance of diarrhea in 15 days. One year later, the patient had resumed his usual weight, and laboratory tests and duodenal biopsies were normal. CONCLUSION: The diagnosis of tropical sprue should be systematically discussed in any malabsorption syndrome with villous atrophy in a patient living or having lived in the tropics.

[Cavernous hemangioma: rare incidentaloma of the adrenal gland]. / L'hémangiome caverneux : un incidentalome rare de la surrénale.

The hemangioma of the adrenal gland is an adrenal gland lesion rare, benign and usually asymptomatic. Discovered incidentally during an abdominal imaging study, it is part of incidentalomas. Imagery is the best to characterise these silent adrenal masses (computed tomography [CT], Magnetic Resonance Imaging [MRI]± Positron Emission Tomography [PET scan] with 18F-FDG). The main risks of the hemangioma are ignorance of malignancy, bleeding and abdominal mass syndrome. The analysis of the literature shows the importance of laparoscopy. A multidisciplinary discussion on this type of lesion appears indispensable both diagnostic and therapeutic.

Trophic interactions between viruses, bacteria and nanoflagellates under various nutrient conditions and simulated climate change.

Population dynamics in the microbial food web are influenced by resource availability and predator/parasitism activities. Climatic changes, such as an increase in temperature and/or UV radiation, can also modify ecological systems in many ways. A series of enclosure experiments was conducted using natural microbial communities from a Mediterranean lagoon to assess the response of microbial communities to top-down control [grazing by heterotrophic nanoflagellates (HNF), viral lysis] and bottom-up control (nutrients) under various simulated climatic conditions (temperature and UV-B radiations). Different biological assemblages were obtained by separating bacteria and viruses from HNF by size fractionation which were then incubated in whirl-Pak bags exposed to an increase of 3°C and 20% UV-B above the control conditions for 96 h. The assemblages were also provided with an inorganic and organic nutrient supply. The data show (i) a clear nutrient limitation of bacterial growth under all simulated climatic conditions in the absence of HNF, (ii) a great impact of HNF grazing on bacteria irrespective of the nutrient conditions and the simulated climatic conditions, (iii) a significant decrease in burst size (BS) (number of intracellular lytic viruses per bacterium) and a significant increase of VBR (virus to bacterium ratio) in the presence of HNF, and (iv) a much larger temperature effect than UV-B radiation effect on the bacterial dynamics. These results show that top-down factors, essentially HNF grazing, control the dynamics of the lagoon bacterioplankton assemblage and that short-term simulated climate changes are only a secondary effect controlling microbial processes.

Association of growth, feeding practices and exercise conditions with the prevalence of Developmental Orthopaedic Disease in limbs of French foals at weaning.

Developmental Orthopaedic Disease (DOD) in limb joints of horses is frequent and is a common cause of pain and lameness. DOD is a multifactorial disease involving genetics, growth, feeding practices and exercise conditions leading to joint injuries. However, there is no clear understanding of the contribution of each factor. The aim of this cohort study was to assess the adjusted effects of breed and gender, growth, feeding practices and exercise conditions on the prevalence of DOD in the limbs of foals at weaning and on the prevalence restricted to osteochondrosis (OC). Twenty-one farms in Normandy, France, were convenience sampled and enrolled in a cohort study. The study sample consisted of 401 foals from 3 breeds, followed from the 8th month of pregnancy of the mares until weaning period at approximately 6 months of age of the foals. Stud farms were regularly visited to collect growth, feeding practices and exercise conditions data. The carpus, the front and hind digits, the hock and the stifle of the foals were X-rayed at the end of follow-up. Foals were categorised as affected or unaffected by DOD or by OC. Two mixed-effects logistic regression models were used to determine risk factors for DOD or OC, adjusting for clustering by farm. Of the 378 foals with complete data, 47% (95% CI: 42-52) were affected by DOD and 36% (95% CI: 31-41) by OC. Risk was significantly increased for DOD and for OC in Warmblood foals compared to Thoroughbreds (OR 2.8; 95% CI: 1.2-6.5 for DOD, OR 2.9; 95% CI: 1.1-7.5 for OC), in foals with a high wither height at 30 days of age, and in foals with a rapid increase of wither height (for DOD only). Exercise conditions such as large area of pasture, some irregular exercise, or some batch changes were also significant risk factors for DOD or OC. No association was found with quantitative feeding practices; it was likely due to the limited variability of horse regimens and to the partial inclusion of nutritional effects in other factors such as breed and growth. In order to reduce the prevalence of DOD or OC in foals, following the growth and reducing nutritional supply in subjects growing fast, limiting the pasture areas offered and providing a regular exercise to foals seems appropriate. To our knowledge, to date the assessment of adjusted effects of potential risk factors for DOD has never been proposed.