The latter allows us to suggest that UA is eliminated from the blood via the gut tissue.An elevated level of serum uric acid—hyperuricemia, is strongly associated with the development of gout and chronic kidney disease (CKD) which is often accompanied by a significantly reduced glomerular filtration rate (GFR). In the present study, we investigated the extra-renal elimination of uric acid via the intestine in a healthy pig model and the effect of oral uricase therapy on plasma uric acid concentrations in pigs with induced hyperuricemia and CKD. The experiment was conducted on eleven, ten-week-old pigs (n = 11). The porcine model of CKD was developed by performing 9/10 nephrectomy surgery on eight pigs. A stable model of hyperuricemia was established in only five of the eight nephrectomized pigs by frequent injections of uric acid (UA) into the jugular vein. All pigs  were operated for implantation of jugular vein catheters and the three healthy pigs also had portal vein catheters inserted. Blood uric acid concentrations were measured spectrophotometrically, using the Uric Acid Assay Kit . The piglets with CKD received orally administered uricase (treatment) and served as their own controls (without uricase supplementation). Oral uricase therapy significantly decreased plasma uric acid concentrations in pigs with CKD, whereas hyperuricemia was observed in the pigs whilst not being treated with uricase. Urinary uric acid excretion was similar during both the treatment and control periods during the first 8 h and 24 h after UA infusions in the CKD pigs. To demonstrate the elimination of UA via the intestine, the healthy pigs were infused with UA into the jugular vein. The blood collected from the jugular vein represents circulating UA concentrations and the blood collected from the portal vein represents the concentration of UA leaving the intestine. The final (after 2 h) concentration of UA was significantly lower in blood collected from the portal vein compared to that collected from the jugular vein (3.34  Most mammals express urate oxidase (uricase) which converts UA to the more soluble allantoin . Howeverturnover . Both inturnover . Due to turnover .Three main pharmacological treatment strategies are currently used in the treatment of HUA. The first treatment strategy is aimed at lowering UA generation and includes the use of xanthine oxidase inhibitors such as allopurinol and febuxostat. However, their effectiveness in lowering serum urate concentrations is limited, since only between 30–60% of patients receiving these therapies reach a plasma UA concentration of < 6 mg/dL . MoreoveThus, the majority of the currently approved therapies for HUA and gout are not recommended for patients with late-stage chronic renal disease. There is therefore a need for alternative, safe and well tolerated therapies which are effective in reducing plasma UA concentrations in patients with late stages of CKD.Based on the peculiarities of urate metabolism and the fact that one third of the daily urate pool is excreted via the small intestine, we were interested in developing an oral enzyme therapy using uricase for patients that have HUA and late stages of CKD. For this purpose we chose to make use of the pig model, as it is widely used in translational research, surgical models, and procedural training in preclinical toxicological testing of pharmaceuticals . HoweverCandida utilis, in decreasing plasma UA concentrations in CKD pigs with induced HUA. We hypothesized that oral uricase therapy may be effective in decreasing serum UA concentrations in CKD pigs, as a result of enhanced intestinal urate elimination.In the present study, we first aimed to develop a reliable pig model for investigation of the extra-renal elimination of UA through the intestine. Secondly, we wanted to investigate the ability of orally administered uricase, from This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The original protocol for this study is available as supporting information; see ad libitum access to feed under similar conditions. During enzyme treatment, the feed was also supplemented with sodium bicarbonate . Water supply was limited and corresponded to an amount of between 10–12% of the pigs’ body weight. During the experimental period food and water intake was measured daily. Pig body weight was measured weekly.The experiment was performed on 11 castrated male pigs ((Swedish Landrace X Yorkshire) x Hampshire) with an average initial body weight of 12 ± 3 kg and an average age of 10 ± 2 weeks at the beginning of the experiment. The pigs were selected from a specific pathogen-free herd . The pigs were divided into two groups–the healthy group (n = 3) and the CKD group (n = 8). Pigs were housed in individual metabolic cages equipped with a dry feeding trough, a drinking nipple and a constant heating lamp (150 W). The metabolic cages allowed for the collection of urine. Pigs were allowed to move freely within their cages and had visual contact with each other. Pigs were maintained on a 12 hour day-night cycle at a constant temperature (22 ± 2°C). Pigs were fed twice daily (2% of their b.w.) with cereal-based feed , with a low calcium (0.09%) and high fructose (20%) concentration. The feed was enriched with inosine  to induce HUA. This amount of feed is comparable to the amount ingested when pigs are given 2 as a carrier gas, at approximately 0.5–1 L/min in a close-circuit respiratory system .Prior to surgery pigs were fasted overnight and pre-medicated with azaperone . The pigs were then anaesthetized with 2-bromo-2-1.1.1-triflouroethane , mixed with air and OTo develop CKD in the pigs, 8 pigs underwent 9/10 nephrectomy surgery. The nephrectomy was achieved by renal tissue infarction. The main left renal artery was completely ligated using sterile silk sutures which resulted in the blockage of blood flow to the entire left kidney. The right renal arterial branches were ligated successively in order to achieve an obstruction to blood flow to 90% of the right kidney. As a result, discoloration of the entire left kidney and 90% of the right kidney was observed, confirming the infarction.In all pigs  the left external jugular vein was catheterized using silicon tubing  with an outer diameter of 1.64 mm and an inner diameter of 0.75 mm. The catheter was exteriorized percutaneously on the dorsal side of the neck.For investigation of the extra-renal elimination of UA through the intestine, the three healthy pigs had an additional catheter inserted into the portal vein, just prior to the entrance to the liver. The portal vein was catheterized using silicon tubing  with an outer diameter of 2.29 mm and an inner diameter of 1.27 mm. Briefly, after localization of the portal vein in the liver hyllus, a double portabac silk  suture (diameter 0.5 cm) was placed on the portal vein wall. A small incision was made in the middle of the portabac suture, then a catheter with fixative cuffs was inserted into the portal vein for 3–4 cm up to the first cuff towards the liver. The portabac suture was then closed over the catheter and the bleeding stopped. Finally, the cuff was gently sutured  to the portal vein wall. Another 3–4 cuffs were placed on the catheter and the catheter was then fixed onto the inside of the abdominal cavity. The catheter was exteriorized using a surgical needle on the right flank of the abdominal wall. A single external cuff protected the catheter from being drawn back into the abdominal cavity. After the surgical procedures, for 3 days following surgery, the pigs received 1.5 mL of Buprenorphine .The pigs were allowed a 2-week recovery period before the experimental period commenced.The current study was comprised of two experiments. In the first experiment the intestinal elimination of UA was investigated in the three healthy pigs (average body weight of 15 ± 3 kg), on two separate occasions with a 1-week interval between the two assessments. Each pig underwent surgery to have a catheter inserted into the jugular vein and one into the portal vein. The blood collected from the jugular vein represents the dynamics of the peripheral circulation, whereas the blood collected from the portal vein represents the dynamics of the intestinal circulation. To reach saturation threshold for UA, the pigs were intravenously infused, via the jugular vein catheter, with a UA suspension  every 30 min during a two hour period; the pigs received four infusions in total. Blood samples were collected from both the jugular and portal veins at baseline (before UA infusions) and thereafter repeatedly at 1 and 2 h following the UA infusion. Urine was collected twice in the 24 h period.Candida utilis  was administered orally to each pig, together with the feed , 1 h prior to and 3 h after beginning the UA infusions. To ensure enzyme stability in the stomach, the pigs received both food and water supplemented with sodium bicarbonate (1%). Uricase from Candida utilis was selected as the lead candidate for potential oral enzyme therapy based on results obtained from previous studies by our lab (unpublished data), as a result of its’ wide pH profile (6–8). The experiment was performed as a two-period, two-sequence crossover study, with the following sequences considered: CT (control-treatment) and TC (treatment-control). The sequences, as well as the treatment/control periods within them were separated by a one-day washout period. Thus, the five CKD pigs were assigned to the CT and then to the TC sequence and served as both the treatment  and control groups (without uricase supplementation), such that each pig served as its own control. Blood samples collected from the jugular vein were collected at baseline (before UA infusions), and then at 2, 4, 6, 8, 10, 12, 16 and 24 h following the first UA infusion. Urine was collected every 8 h during the 24 h period. At the end of experiments the pigs were sedated using azaperone , and euthanized by a single dose of intravenously administered sodium pentobarbiturate (100 mg/kg).The second experiment was performed on five nephrectomized pigs (which were selected from the eight pigs that underwent 9/10 nephrectomy surgery based on elevated plasma creatinine concentrations indicating successful development of CKD), with an average body weight of 16 ± 3 kg at the beginning of the experiment. To induce HUA, pigs were i.v. infused with a UA suspension  every 30 min for an eight hour period–with 16 infusions in total. Uricase from Blood samples were collected into lithium-heparin tubes , that were gently inverted several times after collection and stored on ice before being centrifuged at 3000×g, for 15 min at 4°C. Plasma was collected and the samples were stored at -20°C until further analysis. Plasma UA and creatinine concentrations were analyzed spectrophotometrically using the Uric Acid Assay Kit and QuantiChrom™ Creatinine Assay Kit, respectively , according to the manufacturer’s protocols.2O for UA and 1/20 with 7 mM NaOH for creatinine. Urinary concentrations of UA and creatinine were measured spectrophotometrically using the Uric Acid Assay Kit or QuantiChrom™ Creatinine Assay Kit, respectively , according to the manufacturer’s protocols.To prevent precipitation of UA salts, 1–2 ml of 8 M NaOH was added to the urine collection containers. After each collection, aliquots (about 10 mL) of urine samples were transferred into tubes and stored at –20°C until performing the creatinine assay. UA concentration was measured in fresh urine samples. Before analysis, urine samples were diluted 1/10 with dist Hp < 0.05.All analyses were carried out using Prism, version 5 . Statistical significance in Experiment 1 was evaluated using a two-tailed Student’s t-test followed by a Student-Newman-Keuls multiple comparisons test. For Experiment 2, the total area under the curve (AUC) was calculated for plasma uric acid concentration, urine uric acid concentration (measured at 8 and 24 hours after uric acid infusion), as well as creatinine and urate clearance. Treatment efficacy was assessed based on the within-subject differences between the control and treatment periods with regards to the various outcomes investigated, using the customary formula for an unpaired t-test . A pre-tvs. 3.34 ± 0.52 mg/dL, respectively, p = 0.024). The results obtained suggest that after reaching a certain plasma urate threshold, the elimination of UA via the intestine starts occurring in pigs.Due to the low endogenous plasma urate concentration in pigs (1.2–1.6 mg/dL), in order to reach the saturation threshold for plasma UA the pigs were infused with a UA suspension, via the jugular vein catheter. The 9/10 nephrectomy surgery was performed in pigs to create a model closest to that of a patient with the late stages of CKD. The surgical procedure and postoperative management did not seem to disturb the overall health status and growth of the pigs that underwent surgery, except during the first 6 days after surgery, where no weight gain was observed. Due to the presence of uricase, the nephrectomy did not increase plasma UA concentrations in the pigs . To testA stable porcine model of HUA was achieved by frequent  infusions with a UA suspension (5 mg/kg b.w.) into the jugular vein. This approach resulted in an approximately 2.6-fold increase in plasma UA concentrations during the first 12 h after beginning the UA infusion in CKD pigs, compared to that observed prior to the UA infusions (each pig served as its’ own control).The maximal plasma UA concentration in all pigs was observed 4 h after beginning the infusions . The UA Oral administration of uricase to pigs resulted in an approximately 1.3-fold decrease in plasma UA concentration in CKD pigs, 8 h after beginning the UA infusion, compared to the control period . The mosUrinary UA concentrations are shown in The creatinine and urate clearance were similar in pigs during both sequences, in both study periods and reached 23.8 ± 10.7 and 21.6 ± 18.8 mL/min during the control period, and 19.9 ± 7.0 and 21.3 ± 12.3 mL/min during the treatment period, respectively.Hyperuricemia (HUA) is an abnormally elevated blood UA concentration in humans . When thMost of the drugs targeted at lowering blood UA concentrations are focused on limiting hepatic UA synthesis or enhancing the renal excretion of UA. However, impaired renal function in CKD patients, where HUA results from the under-secretion, rather than overproduction of UA, increases the toxicity of drugs used to treat HUA. Consequently, patients with HUA and CKD are very often left untreated. Therefore, there is a need for the development of alternative therapies that would be effective in lowering serum UA concentrations in patients with CKD and on dialysis for the long-term management of HUA, without gout.To date, a model of HUA has been developed only in rodents. Several reports have demonstrated that in rats HUA can be induced by a high-fat diet and starvation , dietaryCandida utilis, in decreasing plasma UA concentrations in CKD pigs with induced HUA. Even the restriction of kidney function by 90% following 9/10 nephrectomy failed to increase plasma UA concentrations in the pigs used in the current study. However, the nephrectomized pigs demonstrated elevated plasma creatinine concentrations  i.v. infusions of a UA suspension into the peripheral blood, at a dose of 5 mg/kg b.w. We obtained a relatively stable porcine model of mild HUA (around 4 mg/dL) for 8 h . A similIn order to examine whether UA is eliminated via the intestine in pigs, as it is in humans, we compared the plasma UA concentration in the jugular vein blood, representing peripheral circulating UA concentrations with that in the portal vein blood, representing UA concentrations from the intestinal circulation, following the UA infusions. The results obtained clearly demonstrate that UA, after reaching a certain plasma urate threshold is also eliminated via the intestine in pigs, as it is in humans .Uric acid handling is a very complex and dynamic process mediated by multiple specific transporters. In humans, blood urate is freely filtered in the kidney glomerulus . FiltereUnfortunately, there is not much data on the molecular mechanisms of extra-renal excretion of uric acid. It has, however, been shown that intestinal secretion is the major extra-renal elimination pathway of UA . Hosomi Candida utilis was selected as the lead candidate for potential oral enzyme therapy due to its wide pH profile (6–8), which matches that of the small and large intestines, and based on results from previous in vitro studies [Candida utilis. The significant reduction in AUC of plasma UA concentration during the uricase treatment period compared to that observed during the control period —pegloticase. Although, these preparations significantly reduce plasma UA concentrations and act faster than allopurinol, their use in the case of chronic HUA is very limited due to high antigenicity, rapid elimination from the plasma and induction of anti-PEG antibody production [For several decades exogenous uricase has been i.v. administered to test its ability to decrease plasma UA concentrations, but there is a lack of data showing the impact of oral uricase therapy on plasma UA concentrations. Humans and higher primates do not possess uricase and are unable to metabolize UA, and thus, they are at high risk of developing HUA. Clinically approved uricase preparations include uricase from oduction . There hoduction . In contoduction . NeverthUndoubtedly, the present study has certain limitations. The main limitation is the fact that the porcine model doesn’t fully mimic human physiology due to the presence of liver uricase in pigs. A further set of experiments should be performed to clarify the possible mechanisms underlying the results obtained in the present study. Both analysis of UA concentration in the portal vein of nephrectomized pigs and UA concentration in the stool should be performed to fully support the hypothesis that UA is eliminated via the intestine in CKD pigs with HUA. Moreover, while the present study is characterized by a short duration, in order to determine clinical relevance a long-term study needs to be conducted.In summary, the results obtained from the present study in the porcine model suggest the possibility of decreasing plasma UA concentrations in patients with HUA and impaired renal function following oral therapy with microbial uricase, which possibly enhances UA elimination via the intestine. This approach may represent an alternative and safe therapy for the treatment of HUA in patients with attenuated GFR as strongly manifested in CKD.S1 Study ProtocolStudy protocol. Original study protocol.(DOCX)Click here for additional data file.


Cilia mediate an astonishing diversity of processes. Recent advances provide unexpected insights into the regulatory mechanisms of cilium formation, and reveal diverse regulatory inputs that are related to the cell cycle, cytoskeleton, proteostasis, and cilia-mediated signaling itself. Ciliogenesis and cilia maintenance are regulated by reciprocal antagonistic or synergistic influences, often acting in parallel to each other. By receiving parallel inputs, cilia appear to integrate multiple signals into specific outputs and may have functions similar to logic gates of digital systems. Some combinations of input signals appear to impose higher hierarchical control related to the cell cycle. An integrated view of these regulatory inputs will be necessary to understand ciliogenesis and its wider relevance to human biology. Primary cilia integrate complex combinatorial inputs into output functions.Modulatory inputs arise from the cytoskeleton, proteostasis, and ciliary signaling.Cell-cycle inputs lead to irreversible disassembly decisions that override other inputs.The integration of inputs in cilia resembles the behavior of logic gates. Primary cilia are microtubule-based organelles that are formed from a centriolar anchor, known as the basal body, and extend from the apical surface of most mammalian cells . They haWe attempt here to present an integrative viewpoint of pathways that regulate ciliogenesis and the interrelationships between them. A plethora of recent studies suggest that the regulatory inputs into ciliogenesis can be subdivided into four distinct but interrelated cellular processes, each with an intrinsic logic in their function: the cell cycle, structural influences of the cytoskeleton, cellular proteostasis, and signaling processes.In vertebrates, ciliogenesis and cell division are mutually exclusive because the centrioles must be released from the plasma membrane to function in the mitotic apparatus ChlamydomonasHow are ciliogenesis and cilium disassembly coupled to the cell cycle? Because microtubules are in a state of dynamic equilibrium, one obvious mechanism to regulate ciliogenesis is to alter the balance between the assembly and disassembly of axonemal microtubules. Although the mechanistic details are incompletely understood, several centrosomal and ciliary proteins regulate these processes, and their functions frequently depend upon their phosphorylation by cell-cycle kinases. Examples include Polo-like kinase 4 (PLK4) Another member of the kinesin-13 family of microtubule depolymerases, Kif2a, that had previously been implicated in the control of mitotic spindle assembly has also been recently shown to function in ciliary disassembly at the mother centriole Hdac6 mouse mutants do not have any cilia-related phenotypes in vivo redundancy for HDAC6 function. In addition to acetylation, the α- and β-tubulins of the axoneme are also subject to a diverse range of other post-translational modifications . Although this remains to be investigated, other aspects of the tubulin code are also likely to be cell cycle-regulated. This may be the case for the mitotic spindle-associated protein CEP41, which is required for tubulin glutamylation and the organization of axonemal microtubules Aurora A is also implicated in a second mechanism of axonemal disassembly in cells emerging from G0. Aurora A associates with HEF1 Several regulatory circuits link the cell cycle to both the actin cytoskeleton and ciliogenesis . The actCiliogenesis, and presumably ciliary functions, are modulated by the cytoskeleton. Strong evidence supports the role of actin branching in suppressing primary cilia formation. Actin nucleation-promoting proteins such as cortactin, and components of the ARP2/3 complex, which mediates actin branching, inhibit ciliogenesis Actin appears to inhibit ciliogenesis by at least three mechanisms. First, it may have a regulatory role in the transport of cilia-directed vesicles by creating a mechanical barrier to vesicle movement or the membrane remodeling required for ciliogenesis. A role in regulating vesicle transport is supported by observations that actin disassembly results in a transient accumulation of cilia-targeted Smo-positive vesicles at the basal body Chlamydomonas, inhibitors of actin polymerization shorten flagellae and actin mutants cause a decrease in IFT protein recruitment at the cilia base Other pathways also mediate F-actin turnover at the cilia base. Bardet–Biedl syndrome (BBS) proteins and the cytoplasmic dynein light chain, Tctex-1, inhibit and enhance actin polymerization, respectively Cilium interactions with the cytoskeleton are closely related to both planar and apicobasal cell polarity. Cofilin mutants display cilia malpositioned on the apical surface of embryonic node cells, affecting planar cell polarity Viability of the cell depends on the presence of functional proteins in appropriate quantities, maintained through protein synthesis and degradation pathways. There is now compelling evidence that the primary cilium has a role in regulating protein homeostasis by influencing several key protein maintenance mechanisms, including the ubiquitin-proteasome system (UPS), autophagy, and mTOR signaling. The relevance of these three processes to cilia formation and function is discussed below.Chlamydomonas. During flagella disassembly, ubiquitination of flagellar proteins increases, and disassembled proteins such as α-tubulin and polycystin-2/PKD-2 are ubiquitin (Ub)-tagged and transported to the cell body by intraflagellar transport The importance of proteasomal degradation in ciliogenesis first came to light based on studies in Interestingly, UPS-mediated protein degradation is essential for regulating cilia formation and disassembly during the cell cycle A. FollowC. elegans it is dispensable for cilia formation but ensures the correct ciliary targeting of sensory receptors It is important to appreciate that different Ub signals regulate other cellular processes such as endocytosis, for example, that can impinge on ciliogenesis Ciliogenesis is also regulated by the VCP/p97 (hereafter VCP) pathway that separates Ub-labeled proteins from their binding partners and targets them for recycling or degradation by the proteasome. VCP is also involved in autophagosome maturation  complex 1 (TORC1) is a kinase that links amino acid availability to cell growth. It is an important regulator of overall cell metabolism level and a potent inhibitor of autophagy mTOR signaling may also integrate ciliary mechanosensation with global regulation of cell growth. This is supported by observations that the sensation of flow by renal cilia inhibits both mTORC1 activity and cell size through signaling by a ciliary serine/threonine kinase, LKB1 Autophagy is also used for housekeeping purposes such as the degradation of long-lived proteins, elimination of protein aggregates (aggrephagy), and turnover of damaged cellular organelles  C. elegans olfactory neurons change morphology from finger-like to fan-shaped in the absence of particular stimuli, and this has been suggested to enhance their olfactory receptivity An interesting characteristic of the cilium is its ability to adjust morphology in response to environmental conditions. The enlargement of the cilium to enhance its signal detection capacity or, vice versa, resorption to decrease its responsiveness to signals, makes intuitive sense as mechanisms of sensory signal adaptation. Indeed, cilia appear to act as a self-adjusting antenna, and modify their size and shape depending on the strength of signals that they detect. For example, specialized cilia of 2 (PGE2) has been shown to be a positive regulator of ciliogenesis by increasing anterograde intraflagellar transport through its receptor  which localizes to the ciliary membrane, and acts by activating the cAMP signaling cascade 1B serotonin receptors) that localize to the transition zone and mediate ciliogenesis Second messengers and intraflagellar transport play a key role in the adjustment of cilia shape and size in response to extracellular stimuli. Mechanical stimulation of tissue-culture cells decreases cAMP levels and shortens cilia. This is presumably mediated by downregulation of cAMP-dependent kinase signaling and anterograde intraflagellar transport What logic integrates ciliogenesis inputs? The overview of pathways that we have presented above reveals several general themes. First, it appears that interactions of cilia with intracellular processes, such as the cell cycle, autophagy, or cytoskeletal changes involve reciprocal regulatory signaling, presumably providing both positive and negative regulatory feedback loops . For exaIs there a hierarchy to inputs that regulate ciliogenesis? During cellular quiescence, a highly integrated control of multiple inputs may prevent premature and potentially damaging responses to short-term cellular stresses such as random fluctuations in the levels of nutrients, metabolism, cell size, and cytoskeletal deformations. The convergence of multiple inputs in the specific configuration would alter this ‘steady state’ irreversibly, forcing the cell into division. The ensuing disassembly seems to be largely executed by cell-cycle kinases that rapidly phosphorylate centrosomal or ciliary protein substrates, thereby breaking down the very cellular structure that maintains the steady state. Consistent with this logic, the converse also appears to be true: ciliogenesis is induced under conditions of replicative senescence (a stable form of cell-cycle arrest) through the depletion of CP110, the negative regulatory centrosomal protein Outstanding QuestionsWhat is the structural basis of selective permeability at the ciliary gate?How do signaling cascades regulate cilia formation and function?What is the mechanistic basis of ciliary mechanosensation?What mechanisms regulate the function of ciliary proteins that mediate cilia-specific autophagy?What underlies the tissue-specific spatial and temporal differences in ciliogenesis and ciliary function?What other inputs regulate ciliogenesis, and are they organized into a hierarchy?Is there a logic that underlies the integration of these diverse inputs?How can integrated pathways of ciliogenesis, cellular homeostasis, and metabolism be utilized for therapeutic purposes?In conclusion, the cellular ‘antenna’, far from being a passive receiver of input signals, is due for an upgrade to the status of a cellular ‘central processing unit’, and perhaps the main one integrating extracellular signaling with the cell cycle and metabolism. We would argue that an integrated view of the many diverse and complex regulatory inputs and output functions is essential for a full understanding of ciliogenesis and its relevance to human health and disease (see Outstanding Questions). Systems-biology approaches, such as affinity proteomics and reverse-genetics screens of cellular phenotypes or expression profiles, will provide powerful new methodologies to explore the logic circuits and regulatory networks for ciliogenesis.


In recent years, Mechanical Turk (MTurk) has revolutionized social science by providing a way to collect behavioral data with unprecedented speed and efficiency. However, MTurk was not intended to be a research tool, and many common research tasks are difficult and time-consuming to implement as a result. TurkPrime was designed as a research platform that integrates with MTurk and supports tasks that are common to the social and behavioral sciences. Like MTurk, TurkPrime is an Internet-based platform that runs on any browser and does not require any downloads or installation. Tasks that can be implemented with TurkPrime include: excluding participants on the basis of previous participation, longitudinal studies, making changes to a study while it is running, automating the approval process, increasing the speed of data collection, sending bulk e-mails and bonuses, enhancing communication with participants, monitoring dropout and engagement rates, providing enhanced sampling options, and many others. This article describes how TurkPrime saves time and resources, improves data quality, and allows researchers to design and implement studies that were previously very difficult or impossible to carry out on MTurk. TurkPrime is designed as a research tool whose aim is to improve the quality of the crowdsourcing data collection process. Various features have been and continue to be implemented on the basis of feedback from the research community. TurkPrime is a free research platform. For example, finding participants, incentivizing participation, and preventing multiple participation are just some of the common barriers encountered on most crowdsourcing platforms.Access to participants is of central importance to researchers in the social and behavioral sciences. Until the late 1990s, most behavioral researchers had to rely on undergraduate subject pools as their primary source of research participants. University subject pools have a number of limitations, including lack of representativeness . MTurk has in recent years revolutionized behavioral research . For example, Peer et al. . To overcome the current limitations of MTurk, multiple tools have been created to enable researches to accomplish specific tasks  and people who are interested in performing those jobs in return for payment (workers). A task put up by a requester on MTurk is called a human intelligence task (HIT). Requesters can create HITs on the MTurk platform or use MTurk to announce an external HIT. For example, MTurk provides a platform for creating survey questions. A requester may use that built-in MTurk functionality, or instead can create a survey on an external platform such as Qualtrics and post a link to that survey as part of their HIT. We refer to HITs that are completed via an external platform as external HITs, and tasks that utilize the MTurk platform as internal HITs.The MTurk platform was created as a worldwide microtask marketplace. MTurk facilitates monetary transactions between people who need various jobs done  and TurkerNation (http://turkernation.com/), have been created, in part to provide workers with information about the quality of requesters and HITs. Workers commonly provide information on these websites about whether a HIT pays adequately, how long a requester takes to pay for a HIT, and whether a requester rejects HITs unfairly. Requesters should keep track of these websites, because information provided there can affect workers’ willingness to accept a specific requester’s HIT. Additionally, requesters should manage their reputations on these sites, because the information provided there is not always accurate, and can potentially negatively affect participation rate, and thus may compromise the recruitment process.A worker’s approval rating is updated every time the worker finishes a study and is either rejected or approved by a requester. The reciprocal relationship, whereby workers would rate requesters, is not provided by MTurk. However, various websites on which workers leave comments about specific HITs and requesters, such as Turkopticon  have an existing MTurk account, (b) create a TurkPrime account, and (c) link the TurkPrime account to their MTurk account. The TurkPrime website (TurkPrime.com) has a Setup Mechanical Turk tab, which provides the directions for associating a TurkPrime account with an MTurk account. This short process involves providing TurkPrime with a requester’s MTurk account access keys, which are found on their MTurk security credentials page. Once the account is set up, requesters can launch, stop, and make changes to their MTurk HITs directly from TurkPrime.TurkPrime has three primary windows. The Design Survey page see Fig.  is whereTurkPrime studies are launched from the Design Survey page, which can be accessed from TurkPrime’s home page. The Design Survey page is designed to look like the Design HITs page on MTurk. Experienced MTurk users will find that setting up a study on TurkPrime is very similar to doing so on MTurk. For example, requesters will find fields for naming a study, indicating the price, and selecting the quality of workers. The MTurk Design Survey page also contains fields that are not found on MTurk, such as excluding or including participants from prior studies see Fig. .TurkPrime provides a number of ways to limit and grant access to a study for specific workers. In effect, TurkPrime automates the process of assigning qualifications. TurkPrime makes it possible to assign qualifications to participants from multiple studies simply by selecting those studies from a dropdown window or pasting sets of Worker IDs in the Include and Exclude windows.Allowing participants to complete a study multiple times can lead to a reduction in effect sizes  A HIT that is open only to specific workers on the basis of participation in previous studies, (2) matching participants across the phases of a study, and (3) notifying participants that a HIT is available. As we described previously, a HIT that is open only to specific workers can be set up by using the Include control.Because longitudinal studies have multiple phases, there are typically multiple data files that store the results from the different phases. Worker IDs allow data to be matched across phases. TurkPrime facilitates the process of matching Worker IDs across multiple data files by automatically passing each worker’s ID to Qualtrics via an embedded query string in the survey’s URL. Requesters can then automatically insert the Worker ID into data files. In Qualtrics this is easily accomplished by embedding the Worker ID field into the survey flow. As of now, this feature is only available for Qualtrics studies.Opening up a HIT to specific workers differs from a typical HIT in a critical way. Workers who are qualified to take the HIT may not be in front of a computer at the time that a study is launched, and thus may never know that the HIT is available. For this reason, TurkPrime enables a batch e-mail feature for longitudinal studies with which e-mails can be sent to all eligible participants, informing them that the HIT is available, how much it pays, and any other information that a requester may wish to include to incentivize the workers to participate in the study. The e-mail automatically includes the HIT link.TurkPrime has a separate Worker Groups window that enables the creation of panels, which can be considered to be a special case of longitudinal studies. Panel groups are collections of workers who share common characteristics. For example, these can be political groups such as Republicans or Democrats. Groups of Worker IDs can be entered into the Groups window and given a name. Once a group is created, it becomes visible in the Groups dropdown menu on the Design Survey page. A study can then be created that is open only to the participants of these groups. Worker group studies can be especially effective when requesters have accumulated a large number of workers that can then be utilized as a requester’s own subject pool . These platforms vary widely in the range of tools that they offer and the GUIs that they provide. One platform that offers a wide range of tools is psiTurk. Like TurkPrime, psiTurk offers the abilities to exclude participants, offer bonuses, and make automatic payments, among many other features. psiTurk is open-source platform for designing flexible online studies  for all transmitted data. All data access is blocked except for explicitly white-listed IP addresses, in addition to being secured with user passwords. Furthermore, MTurk data, including Access Key ID and Secret Access Key, are encrypted with AES-256 encryption, the standard adopted by the National Institute of Standards and Technology.To summarize, TurkPrime was created as an extension of Amazon Mechanical Turk to optimize MTurk functionality for the needs of researchers. MTurk is a powerful crowdsourcing environment that provides substantial advantages over other online subject recruitment platforms. Among these are fast and easy access to thousands of research participants, a payment mechanism to incentivize participation, a way to prevent multiple participation by the same individual, and a high level of confidentiality. In recent years, MTurk has become the most popular crowdsourcing research platform, with close to 1,000 peer-reviewed papers published from its data per year.The vast power of the MTurk API, however, has been considerably underutilized by most researchers. TurkPrime offers a programming-free user interface that enables researchers to harness the full power of MTurk. TurkPrime enables researchers to employ research designs, including longitudinal, time-series, and panel studies, that would otherwise be difficult and time-consuming, and that would require substantial API programming to implement. TurkPrime enhances the ease with which requesters can communicate with workers, including sending batch e-mail notifications and bonus payments; provides a flexible way to break up studies over time; gives users complete control over launched HITs; automates the approval/rejection process; provides Qualtrics integration; and allows for the creation of custom worker groups for inclusion and exclusion across studies. Worker groups have the additional benefit that requesters can notify workers who are not monitoring MTurk at the time of study launch. This makes it possible for requesters who have accumulated large worker pools to reach more workers than they would be able to by launching a study on MTurk alone.TurkPrime was created as a dynamic environment that is responsive to the needs of researchers. As such, new features have been recommended by the research community and are being added in an ongoing effort to enhance the quality and usability of online research.


Tissue polarimetric imaging measures Mueller matrices of tissues or Stokes vectors of the emergent light from tissues  over a field of view, and has demonstrated utility in a number of surgical and diagnostic applications. Here we introduce a compact complete Stokes polarimetric imager that can work for multiple wavelength bands with a frame-rate suitable for real-time applications. The imager was validated with standard polarizing components, and then employed as a polarization state analyzer of a Mueller imaging polarimeter and a standalone Stokes imaging polarimeter respectively to image the process of dehydration of bovine tendon tissue. The results obtained in this work suggested that the polarization properties of the samples rich of collagen fibres can change with the degree of dehydration, and therefore, dehydration of the samples prepared for polarimetric imaging (e.g. polarimetric microscopy) should be carefully controlled. Polarization imaging has aroused interest in the field of tissue imaging since it can be a powerful tool to reveal morphological, structural and compositional information of tissue. Tissue polarimetric imaging measures  Mueller matrices of biological tissues or  Stokes vector of the emergent light from tissues  over a field of view. Tissue Mueller polarimetric imaging has demonstrated utility in a number of surgical and diagnostic applications , like cain vivo polarimetric imaging. As a fast PSA, it can also accelerate the acquisition speed and pave the way for fast Mueller polarimetric imaging.A Stokes polarimetric imager is a core device for Stokes polarimetric imaging techniques, and is also an essential component of a Mueller imaging polarimeter known as the polarization state analyzer (PSA for short). A real time, lightweight and compact Stokes polarimetric imager that can work under multiple wavelength-bands in the visible range is desirable since it would be easier to integrate such an imager with endoscopes and microscopes to do Stokes polarimetric imagers can be constructed with mechanically rotating waveplates or linear polarizers. A polarimeter with mechanically moving parts has disadvantages like relatively slow speed, vibration, shorter life etc., so mechanically moving parts are avoided where possible when polarimetric imagers are designed. Stokes imagers with two temperature controlled liquid crystal viable retarders (LCVRs) working together to time-sequentially generate four pairs of phase retardance to reconstruct one Stokes image have been demonstrated . At leasThe integration of a pixelated micro-linear-polarizer array (LPA) directly onto an image sensor has become achievable nowadays to measure the polarization states in a single snapshot. However, the vast majority of this kind of devices are only sensitive to linear polarization. There are only several exceptions which were based on customized pixelated micro-elliptical-polarizer arrays with liquid-crystals and wire grid linear polarizers –20. The Here we demonstrate a compact complete Stokes polarimetric imager based on a LPA camera and a LCVR. The imager can work for multiple wavelength bands with the frame rate suitable for real time applications. It has been validated with standard polarizing components, and was then employed as the PSA of a Mueller imaging polarimeter and a standalone Stokes imaging polarimeter respectively to image the process of dehydration and associated optical clearing of bovine tendon tissue.S is a four-element vector used to characterize polarization state of light. According to its definition, S can be determined by a set of radiometric intensity measurements using “horizontal” (HI), “vertical” (VI), + 45° (I45), −45° (I-45) linear, as well as left (LI) and right (RI) circular polarization analysers.DOP) of light refers to the intensity of the polarized portion of the light out of its total intensity, and can be calculated using the following equation,DOP is 1 for fully polarized light, 0 for unpolarized light, and in the range 0-1 for partially polarized light.A Stokes vector M can transform an input Stokes vector Sin to an output Stokes vector Sout, expressed as The polarization state of light may be transformed during polarized light-sample interactions, depending on the polarization properties of that sample. The mathematical description of the transformation is a Mueller matrix defined in the Stokes vector space. A Mueller matrix δ and θ are the magnitude of its linear retardance and fast axis orientation respectively. The LCVR was modulated between two retardance states in the temporal domain, and one image was acquired by the LPA camera per retardance state, so that the DRM of the Stokes polarimetric imager in S can thus be obtained according to the following equation,−1 and –T denote Moore–Penrose pseudoinverse and matrix transpose respectively, and In is a column vector containing the intensity readings from each pixel of a super-pixel under the two retardance states δ1 and δ2 respectively.As displayed in Fig. 1(a)LCVRs generally produce high retardance with low driving voltage and low retardance with high driving voltage. The switching time from a high to a low retardance state is less than 1 ms, but it typically takes much longer from a low to a high retardance state, as shown in the diagram in The photo of this Stokes polarimetric imager was shown in DRM of the Stokes polarimetric imager in order to minimize the error propagating from the measured radiometric vector In to the reconstructed Stokes vector S in DRM is a function of θ, δ1 and δ2. The optimization can be conducted by finding the optimal θ, δ1 and δ2 so that the 2-norm condition number of the DRM reaches its minimum  given in  for 628 nm band with the standard deviation 0, 0.015, 0.005, 0.019 for S0,1,2,3.The Stokes polarimetric imager was validated using a linear polarizer rotated from 0° to 180° in 10° steps for a 546 nm and 628 nm narrowband light source. The Stokes polarimetric images were reconstructed according to Fig. 4As demonstrated in DRM−1 matrix and a 8 × 1 In matrix according to S1 image in S1 and S2 were expected.Reconstruction of Stokes polarimetric images was based on Visualization 1. The video frame rate reached 16 frames/second with 15 ms exposure time for each individual image. The frame rate can be further increased by reducing exposure time or enabling the multiple-tap function of the camera . It has been illustrated in this experiment that the Stokes polarimetric imager can be used to determine the Stokes vector images in real-time under multiple wavelength-bands. This imager can easily work with other wavelength-bands by simply changing the drive voltages for optimization purposes.A Stokes polarimetric video was also generated when this sample was moving, and was shown in ex vivo bovine tendon) at a reasonable frame-rate. The sample was exposed in the air and underwent evaporation, resulting in dehydration over a period of about 16 minutes. Mueller polarimetric images were acquired every minute to monitor the change of polarization properties of the sample, and a video file that contains all of the acquired Mueller images is shown in Visualization 2. The Mueller polarimetric images were then decomposed based on Lu-Chipman’s polar decomposition method, and the magnitude of diattenuation, total depolarization, the magnitude of retardance and fast axis orientation images were then reconstructed , and the DOP images of the emergent Stokes measurements denoted by DOPs were displayed in SDOP images demonstrate a similarity to the diattenuation image, and the change of SDOP with dehydration was highly consistent with diattenuation.The three fundamental polarization properties are diattenuation, retardance and depolarization  and a Muex vivo bovine tendon). In this experiment, unpolarized light (the 546 nm band) was used to illuminate the sample, and the Stokes polarimetric imager was used to image the sample in a transmission mode. The sample underwent evaporation in the air like the last sample, resulting in dehydration that lasted for 14 minutes. The Stokes polarimetric images were acquired every minute to monitor the change of polarization property of the sample. The results obtained at 0 min, 7 min and 14 min are demonstrated in Fig. 6(a)-6(c)S1 images. The DOPs images of the sample representing tissue diattenuation dropped as with tissue dehydration  should be carefully controlled to guarantee consistency in inter- and intra- sample inspection. It is expected that this imager could enable The authors declare that there are no conflicts of interest related to this article.


The Fas/FasL system is known, first and foremost, as a potent apoptosis activator. While its proapoptotic features have been studied extensively, evidence that the Fas/FasL system can elicit non-death signals has also accumulated. These non-death signals can promote survival, proliferation, migration, and invasion of cells. The key molecular mechanism that determines the shift from cell death to non-death signals had remained unclear until the recent identification of the tyrosine phosphorylation in the death domain of Fas as the reversible signaling switch. In this review, we present the connection between the recent findings regarding the control of Fas multi-signals and the context-dependent signaling choices. This information can help explain variable roles of Fas signaling pathway in different pathologies. Fas TNFRSF6/CD95) belongs to the tumor necrosis factor receptor superfamily. When bound to Fas ligand (FasL) or agonistic antibodies, Fas can recruit Fas-associated death domain-containing protein (FADD), procaspase-8, procaspase-10, and cellular FLICE inhibitory proteins (c-FLIPs). This leads to the formation of the death-inducing signaling complex (DISC), the caspase cascade and ultimately apoptosis . Apoptos/CD95 belCurrent Fas-targeting therapies aim to activate or inhibit Fas signaling , 14. HowPhysiologically, the presence of different FasL forms is an important extrinsic factor that can influence Fas signaling modes. Membrane-bound FasL is essential for activating Fas-mediated apoptosis and thus instrumental in the safeguard against autoimmunity and cancer. Meanwhile, excess soluble FasL (sFasL) may promote autoimmunity and tumorigenesis through non-apoptotic activities . HoweverFas possesses the protein-interacting domain, death domain (DD) , 16. FasUnbound human Fas (hFas) DD comprises six α-helices  Figure A. Fas DDWhile structural studies shed light into the DISC formation, an important question remained: “What gives the cue for Fas DD to form the DISC or a non-death-inducing signaling complex (NDISC)?” Because tyrosine Y232 and Y291 in hFas DD are phosphorylable , tyrosinOur recent study based on the analysis of evolution-guided Fas pY proxies and site-specific pY detection has revealed that DD tyrosine phosphorylation of hFas is the reversible antiapoptotic/pro-survival multi-signaling switch, namely, the DD tyrosine phosphorylation turns off the proapoptotic signal and turns on the pro-survival signals by dominantly inhibiting the DISC formation and apoptosis Figure B while pFAS allele and that the signal transition from non-apoptotic to apoptotic signaling occurs when the Fas signal strength, exhibited by wild-type Fas protein, exceeds a threshold. This opinion is based on two reasons: (1) a mutated FAS allele that causes the loss of apoptotic function is often considered completely non-functional and (2) when Fas mutations are detected, tumors rarely have the loss of heterozygosity  to reach its high threshold, while the threshold for the non-apoptotic signal is so low that it is attainable with one wild-type FAS allele , including Src, Yes, Fyn, Blk, Yrk, Fgr, Hck, Lck, and Lyn, are protein tyrosine kinases that are preferentially expressed in different tissues , 31. DatStudies of hFas in human colorectal cancer (CRC) cells have shown that Src and Yes play an important antiapoptotic and pro-survival roles in hFas signaling by phosphorylating hFas at Y232 and Y291 . The phoA point to keep in mind is the context under consideration. The roles of SFKs in Fas signaling and even the identity of the SFKs involved may differ appreciably in different tissues, disorders, or disease stages since expression profiles of kinases can vary significantly from one setting to another. For instance, while Src and Yes are key regulators of hFas phosphorylation in some solid tumors, this may not hold true for some hematopoietic malignancies where other oncogenic SFKs, such as Lck or Fgr are prominently present. Additionally, divergence in terms of regulatory specificity exists among model systems. For example, a non-conservative tyrosine phosphorylation site in Fas DD among primates and rodents  suggestsSrc homology domain 2 (SH2)-containing tyrosine phosphatase-1 (SHP-1), a protein tyrosine phosphatase, is predominantly present in hematopoietic cells and to a lesser extent in other cell types, including epithelial cells –48. HumaNotably, the rodent models demonstrate that the roles of SHP-1 depend on contexts, including tissues, diseases, and species. For example, Fas-mediated apoptosis was defective in the lymphoid organs but not in hepatocytes and thymocytes from SHP-1-deficient mice , 53. In Besides its cognate ligand and agonistic antibodies, other activators, including anticancer drugs and cytokines, can also activate Fas signaling –59. As wThe involvement of tyrosine phosphorylation in Fas signaling has been well appreciated. In fact, the pY-based survival/apoptotic switch system also applies to other actors in Fas signaling network, with an important example being Caspase-8.Human Caspase-8 (hCaspase-8) has at least three tyrosine phosphorylation sites. Phosphorylation of Y310, Y397, and/or Y465  suppresses Fas-mediated apoptosis –72. AddiNotably, Caspase-8 in rodents, including mouse, rat, guinea pig, and Chinese hamster lacks tyrosines at the positions equivalent to Y310 and Y380 in hCaspase-8. In mouse and guinea pig, it also lacks the tyrosine at the position equivalent to Y465 in hCaspase-8. Therefore, Caspase-8 multi-signaling regulation in rodents is likely to differ significantly from that of human, particularly regarding the dependence on tyrosine phosphorylation switch. Similar to Fas, this highlights the importance of careful consideration when applying the observed regulation and functions of Caspase-8 from one species to another. The lack of a tyrosine at the position equivalent to Y310 (SHP-1-binding site) of hCaspase-8 in rodent SHP-1 coincides with the observation that it participates in Fas signaling differently from human SHP-1. Thus, it is quite probable that, in different species, distinct sets of kinases and phosphatases regulate Fas multimode signaling system. This observation, along with other species-based variations in immune-related signaling , 77, serThat the SFK/SHP-1 phosphorylation-dephosphorylation mechanism applies to both Fas and Caspase-8, the main DISC components, based on information from human cells, suggests at least one common “survival-ON/death-OFF” switch system in the Fas signaling network and can help explain some observation of Fas-mediated outcome in human pathologies. For instance, hyperactivated Src and/or downregulation of SHP-1, such as in some cancers, may favor Fas DD and Caspase-8 tyrosine phosphorylation and thus the antiapoptotic/survival mode of Fas signaling.FAS gene but also on an intricate signaling network involving not only different forms of its ligand but also direct and indirect regulators, which varies according to genetic background. In the clinical aspect, parameters governing Fas multi-signaling may be divided into two main groups: Fas mutation-influenced parameters (MIPs) and regulator-influenced parameters (RIPs)  Figure D. FollowAutoimmune lymphoproliferative syndrome (ALPS) is often affected by MIPs since Fas mutations, mostly in DD, are the common cause of the disease . Some FaFAS gene polymorphisms and mRNA editing. Fas gene polymorphisms have been associated with susceptibility to SLE  illustrates multi-faceted MIPs and RIPs that involve both the downregulation of apoptosis and the upregulation of migration mediated by Fas. MIPs in SLE include y to SLE , althougpatients . While tpatients . Meanwhipatients , can alspatients . Elevatepatients , 95 are patients . The sFapatients . MeanwhiThe progression of GBM, where Fas mutations are uncommon , involveA hyper-apoptotic signaling is significant in myelodysplastic syndromes (MDS) , where ROverall, the findings discussed here emphasize the importance of examining Fas multi-signaling while considering different contexts that can influence Fas signaling switch. A context-oriented understanding of Fas multi-signaling will allow the efficient design of Fas-related therapeutic strategies.All authors listed have made substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This study aimed to assess the reliability of immediate UA measurement with capillary blood samples obtained by fingertip puncture with the HumaSensplus meter in the clinic and the routine plasma UA method in the biochemistry laboratory of 238 consenting diabetic patients. HumaSensplus capillary and routine plasma UA measurements were compared by linear regression, Bland-Altman plots, intraclass correlation coefficient (ICC), and Lin’s concordance coefficient. Values outside the dynamic range of the meter, low (LO) or high (HI), were analyzed separately. The best capillary UA thresholds for detecting hyperuricemia were determined by receiver operating characteristic (ROC) curves. The impact of potential confounding factors  was assessed. Capillary and routine plasma UA levels were compared to reference plasma UA measurements by liquid chromatography-mass spectrometry (LC-MS) for a subgroup of 67 patients.UA levels were measured using both the HumaSensIn total, 205 patients had capillary and routine plasma UA measurements available. ICC was 0.90  0.87–0.92), Lin’s coefficient was 0.91 (0.88–0.93), and the Bland-Altman plot showed good agreement over all tested values. Overall, 17 patients showed values outside the dynamic range. LO values were concordant with plasma values, but HI values were considered uninterpretable. Capillary UA thresholds of 299 and 340 μmol/l gave the best results for detecting hyperuricemia . No significant confounding factor was found among those tested, except for hematocrit; however, this had a negligible influence on the assay reliability. When capillary and routine plasma results were discordant, comparison with LC-MS measurements showed that plasma measurements had better concordance: capillary UA, ICC 0.84 (95% CI 0.75–0.90), Lin’s coefficient 0.84 (0.77–0.91); plasma UA, ICC 0.96 (0.94–0.98), Lin’s coefficient 0.96 (0.94–0.98).plus meter were reasonably comparable with those of the laboratory assay. The meter is easy to use and may be useful in the clinic and in epidemiologic studies.UA measurements with the HumaSens Gout is the most common form of inflammatory arthritis and its incidence has increased over the past decade, with the prevalence now exceeding 1% in most developed countries . It can Gout results from chronic monosodium urate (MSU) crystal deposition in and around the joints caused by longstanding hyperuricemia . Thus, aUA is presently routinely measured by a uricase-based laboratory assay on plasma samples obtained after venous puncture . An accuplus device demonstrated good usability and assay precision, with a coefficient of variation from 4.5% to 8% in five patients who had multiple measurements [R2) was 0.757 in comparison with liquid chromatography-mass spectrometry (LC-MS), the reference method for UA quantification [plus point-of-care meter, to determine the best capillary UA thresholds that could be considered predictive of a plasma UA of 360 and 300 μmol/l, and to identify potential confounding factors.Point-of-care UA meters recently commercialized in Europe allow for immediate UA measurement in capillary blood samples obtained by a fingertip puncture, a method as simple as the commonly used capillary glycemia measurement. A comparison of several of these meters concluded that the HumaSensurements . The coefication , and 0.8This was a monocentric cross-sectional study performed at a diabetology day-care center of a French hospital in Paris. The study was approved by the local ethics committee.The patients included in the study were all consecutively seen in the diabetology day-care center between January 2016 and March 2016. All were diabetic, 18 years or older, and were able to understand the requirements and purpose of the study. Patients with gout were not excluded. Patients received a written information sheet with details on the study. All gave informed consent before any study procedure. A subset of patients agreed to sign a specific informed consent for LC-MS measurement. A venous puncture for standard blood biochemistry, which included plasma UA, and a fingertip puncture for capillary glycemia measurement were routinely scheduled in the diabetology day-care center. No supplementary puncture was necessary for this study.plus UA meter (European Conformity marked and approved for EU market use only) and compared to a standard UA measurement in the hospital biochemistry laboratory that used a venous sample taken and prepared as Li-Heparin plasma and an Abbott uricase automated colorimetric assay. LC-MS was performed on frozen Li-Heparin samples in AstraZeneca’s Protein Biomarkers Laboratory . Medical information  were retrieved from medical files.Capillary UA was measured using the HumaSensplus meter has a dynamic measurement range of 180–1190 μmol/l (3.0 to 20.0 mg/dl). When the meter read LO or HI, indicating that the individual’s UA value was outside the test range, these values were individually compared to corresponding plasma measurements.The HumaSenst test. Differences between methods were calculated for each participant, and the distribution was graphically displayed with kernel density estimation (a nonparametric estimation of density). Agreement between methods was assessed by the intraclass correlation coefficient (ICC) and the Lin’s concordance criterion [The statistical analysis involved all patients with available capillary and plasma UA measurements. Results are expressed as mean ± SD. Capillary and plasma UA measurements were compared by paired Student riterion . When meriterion . In addiplus capillary UA, plasma UA determined by uricase automated colorimetric assay, and plasma UA determined by LC-MS. Differences between methods were calculated. ICC and Lin’s concordance coefficient were calculated between each pair of methods. Results for capillary UA and plasma uricase UA were plotted against plasma LC-MS UA results. Concordance between capillary UA and LC-MS UA data and between plasma uricase UA and LC-MS UA data were graphically assessed by the Bland-Altman method. The analysis involved the use of SAS 9.4 .Additional analyses were performed in the subset of patients with measurements of HumaSensn = 16) or with capillary UA outside the HumaSensplus dynamic range (n = 17). Patients’ demographic characteristics are presented in Table n = 134; 65.4%), mean age was 58.1 ± 13.4 years, 151/201 (75.1%) had type 2 diabetes, 140 (68.6%) had hypercholesterolemia, and 120 (58.8%) were hypertensive.Two hundred and thirty-eight patients were included in the study. The statistical analysis involved 205 patients after excluding 33 patients with missing capillary or plasma UA measurement n = 6 or withNineteen capillary samples were marked LO by the meter (capillary UA < 180 μmol/l). A second measurement was performed with the meter, and values were obtained for four patients: 180, 180, 270, and 400 μmol/l. Those values were included in the statistical analysis; the other 15 patients were excluded and analyzed separately. Among these 15 patients, 12 had UA values below 180 μmol/l according to plasma measurement, and three had values above: 189, 206, and 428 μmol/l. Two samples were marked HI by the meter (capillary UA > 1190 μmol/l). These were excluded from the statistical analysis and were measured at 303 and 213 μmol/l by the laboratory uricase assay.p = 0.0010). The linear regression formula was capillary UA = 39.21 + 0.85 × plasma UA  and mean plasma UA was 309 ± 90 μmol/l (5.19 ± 1.51 mg/dl). The mean difference between capillary blood and plasma UA was −8.6 ± 36.9 μmol/l  and Lin’s concordance coefficient was 0.91 0.88–0.93) , the best capillary UA threshold according to the ROC curve was 340 μmol/l, with sensitivity and specificity 89% ; however, improvement of the determination coefficient when correcting for hematocrit was negligible, from 0.83 to 0.86. The main types of medication patients received are shown in Table p = 0.27).Among potential confounding factors, only hematocrit significantly influenced capillary UA measurements  Table . Agreemeplus meter was easy to use and gave results that were reasonably comparable to those of the uricase laboratory assay within the meter’s dynamic range. Accuracy and agreement with plasma UA measures were good.In assessing the reliability of immediate UA measurement of capillary blood samples obtained by fingertip puncture, the HumaSensA limitation of this study is that the data were obtained from diabetic patients and not gout patients for ethical reasons and ease of implementation since no supplementary puncture was necessary in this population. Thus, the results cannot be generalized to gout patients and therefore we plan to confirm these results in a large population of gout patients.plus capillary blood UA measurement among demographic and biological parameters tested or treatments. However, the potential influence of specific gout treatments, too infrequent in this population, could not be assessed. Hematocrit can influence capillary UA measurements by modifying blood viscosity since capillary UA is measured on whole blood. The manufacturer recommends the use of the HumaSensplus meter for a hematocrit range of 30% to 55%. The results of this study confirmed the influence of hematocrit on capillary UA measurements, but it had no real effect on the assay reliability.We found no clinically significant confounding factor for HumaSensIn some cases, discordance was observed between capillary and plasma UA measurement. The plasma uricase-based method gave better agreement with the reference method (LC-MS) than capillary UA measurement, especially in those cases. Factors that could affect the capillary UA meter results include tissue perfusion/peripheral blood flow, acid-base status, and temperature. Additionally, according to the manufacturer’s instructions, a cell phone being used in the vicinity could produce potential electronic signal interference. Operator error could also play a role; thus, a second measurement could be considered.plus range, are very rare in gout patients.Regarding values outside the meter range, most of the confirmed LO values were concordant with plasma UA; in contrast, the two HI values obtained were not concordant and can be considered uninterpretable until we have further data. A repeated test should also be considered in these cases. However, UA values > 1190 μmol/l, the upper limit of the HumaSensFor the target plasma UA levels of 360 and 300 μmol/l, the capillary blood UA thresholds of 340 and 299 μmol/l, respectively, seemed appropriate to discriminate patients with hyperuricemia, the objective of UA measurement in gout.plus devices could be recalibrated by using the linear regression formula capillary UA = 39.21 + 0.85 × plasma UA.To improve the results and avoid using capillary UA target threshold values different from the plasma UA threshold commonly used, HumaSensplus meter achieved 93.2% values within a deviation of 20% as compared with plasma uricase measurements.Point-of-care glycemia testing meters have been developed and used for many years, and from 2003 the ISO15197 guideline has established a 20% (% deviation) accuracy requirement in comparison to the reference method to achieve regulatory approval (95% of the measurements must be in the acceptance range) . RecentlGood adherence to urate-lowering therapy ranges from only 18% to 44% in gout patients , and 20%Point-of-care testing and self-management have shown benefits in other clinical areas. Daily self-monitoring of blood glucose has been found to be useful for maintaining glycemic control in noninsulin-treated type 2 diabetes , and immMoreover, immediate measurement before pegloticase infusion would facilitate the use of the drug since reinfusions should not be performed in patients with high UA . Point-oplus meters were easy to use and gave results that were reasonably comparable to those of the uricase laboratory assay. HumaSensplus-based capillary blood UA measurements can be useful in the clinic and in epidemiologic studies. Further studies are needed to determine the performance of the meter in gout patients and whether frequent UA self-measurement can improve patient adherence to urate-lowering therapies and achievement of target UA levels, especially when combined with educational programs and/or mobile application use.For reliable, immediate UA measurement in capillary blood samples obtained by fingertip puncture, HumaSens


Multiple quantum modes in correlated fields are essential for future quantum information processing and quantum computing. Here we report the generation of multi-mode phenomenon through parametric amplified four- and six-wave mixing processes in a rubidium atomic ensemble. The multi-mode properties in both frequency and spatial domains are studied. On one hand, the multi-mode behavior is dominantly controlled by the intensity of external dressing effect, or nonlinear phase shift through internal dressing effect, in frequency domain; on the other hand, the multi-mode behavior is visually demonstrated from the images of the biphoton fields directly, in spatial domain. Besides, the correlation of the two output fields is also demonstrated in both domains. Our approach supports efficient applications for scalable quantum correlated imaging. Multi-mode is a fundamental characteristic for quantum information technologies13578917182021In this paper, we deepen the multi-mode research in a system coexisting of four- and six-wave mixing (SWM)23E1, the three level “double-Λ” rubidium (Rb) atomic configuration is formed, involving two hyperfine ground states of 5S1/2 [F = 2 (|0〉) and F = 3 (|1〉)] and an excited state 5P3/2 (|2〉). The spatial beams alignment and energy-level diagram are shown in E1  is set as 780.23 nm with power up to 100 mW and the temperature of media set as around 145 °C, a spontaneous parametricωs) and anti-Stokes (ωas) fields. The two generated fields are symmetric to the axis of E1 with the angle of 0.26°, satisfying the phase matching conditions ks = 2k1 − kas and kas = 2k1 − ks. Then the weak probe field Ep  intersects with E1 inside the Rb cell with the same angle of 0.26°. When the frequency of Ep is the same as Stokes or anti-Stokes signal (ωp = ωs or ωp = ωas), Ep could be treated as injected into Stokes or anti-Stokes field, while the other non-injected port is termed as the conjugate channel. Thus the parametric amplifiedAt the beginning, when we only apply a strong pumping field Gij = μijEij/ħ (j = s and as) is the Rabi frequency, Γij = (Γi + Γj)/2 is the decoherence rate between |i〉 and |j〉; Δi is the detuning between the resonant transition frequency Ωi and the laser frequency ωi of Ei, denoted as ∆i = Ωi − ωi. And we define 1 − Δs = 0 when probe beam has the same frequency with Stokes field, i.e. Δp = Δs, where P = Δas.where E2  at the opposite direction of E1 as injection into Stokes field, shown in E2 acts as a dressing field connecting the transition 5P3/2 (|2〉) to 5D5/2 (|3〉). The presence of E2 will lead to an electromagnetically induced absorption (EIA)ks = 2k1 − kas + k2 − k2 and kas = 2k1 − ks − k2 + k2, shown in Fig. 1 is splited by E1. Multiple split-peaks are direct evidences of multi-mode in frequency domain. Specifically, by increasing the diameter of E1, the intensities of both Stokes and anti-Stokes PA-FWM gain peaks first grow to maxima then decrease. Moreover, there exists the competition between the left and right portion of the signal: in We begin with the case of frequency multi-mode through FWM. Here only pump beam E1, whose diameter is associated with the nonlinear phase modulationHere the splitting is caused by the internal-dressing effect of δ is introduced. Besides, an additional nonlinear phase shift factor eiΦ is introducedE1, compared with δs = 0 and δas = 0 correspond the suppression conditions Δs = Δ1 and 1 = −1.1 × 2π GHz, E1-induced splitting energy levels, which means the switch of enhancement and suppression. And when the diameter changes from 0.8 mm to 3.2 mm, Ф goes through 1.625 π. The normalized theoretical simulations , the EIA dip moves with ∆2 in ks = 2k1 − kas + k2 − k2 and energy conservation in Stokes and anti-Stokes fields, the enhancement of anti-Stokes signal leads to the Stokes peak in conjugate channel , δs+(δas+) to δs0(δas0) are stronger, which means the Stokes (anti-Stokes) signal has larger probability to appear at δs−(δas−) or δs+(δas+). The simulations suggest that stronger dressing effect will result in larger probability of phase mismatch and multi-mode. In short, the multi-mode phenomenon of FWM is caused by internal dressing effect, while in case of SWM, it is subjected to strong dressing effect of E2. Besides, SWM has better potential to achieve more modes than FWM.Theoretically, we also conduct Stokes and anti-Stokes simulations versus phase mismatch p = ∆s so the probe field is injected into Stokes field (while the conjugate channel means anti-Stokes field), and change Δ1 from −0.9 × 2π GHz to 0.3 × 2π GHz. Theoretically, with the phase-mismatch δ in frequency, we have ∆k = δ((1/vas)kas0 −  (1/vs)ks0) from Ep is not injected, each pair of correlated Stokes and anti-Stokes photons could distribute evenly between the ideal Stokes and anti-Stokes cones due to the perfect symmetryEp in our experiments, the intensities in probe and conjugate fields are amplified with multiples of G and (G−1)After discussion about multi-mode in frequency domain, in 1 in 1; and the fields intensities are strengthened. Specifically, the shape of the spatial mode varies with ∆1. In 1 = −0.9 × 2π GHz) there is only one small bright spot (denoted on figures by circle), later three more spots  appear subsequently as Δ1 gradually changes to −0.4 × 2π GHz. As the four spots have different responses for ∆1, it testimonies that the four spots represent different spatial modes. So instead of using noise figure (NF)1 in 4Experimentally, the detailed spatial images of both probe (Stokes) and conjugate (anti-Stokes) fields are shown. Initially, we focus on the evolutions of Stokes and anti-Stokes spatial images versus ∆E1, the normalized simulations of Stokes and anti-Stokes signals versus δ at different ∆1 are shown as δs and δas, respectively. Firstly, the absolute values of δas are generally much larger than δs, so it is reasonable that there are more spatial modes in anti-Stokes than Stokes field. The deduction fits well with the appearance that there are more light spots in the images of anti-Stokes (/Aone). The two beams are of orthogonal polarization, intersecting at the rubidium cell, which couple the three-level “double-Λ” PA-FWM configuration. If dressing beam E2  is injected opposite to E1, the four-level “inverted-Y” PA-SWM process is formed. The 1 cm rubidium cell is of natural abundance, including both 85Rb and 87Rb, and wrapped by μ-metal and heated by a heater tape. Both the probe and conjugate signals are detected by a balanced photodetector (105 V/A) with the diameter of around 0.5 cm, or shot by a CCD.The pump beam n2, will be added into the refractive index at the intersection point. Here we have Gc is the Rabi frequency of conjugate field, and χ(3) is the third-order nonlinear susceptibilities. The nonlinear phase shift Ф is introduced as kp is the length of probe field wave vector.The so-called cross-Kerr effect occurs when a weak probe beam interacts with a strong pump beam, then the spatial orientation of probe beam will be shifted, and a nonlinear term, How to cite this article: Zhu, D. et al. Multi-mode of Four and Six Wave Parametric Amplified Process. Sci. Rep.7, 43689; doi: 10.1038/srep43689 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


While recent studies have indicated that fertility has remained high in Uganda, no systematic attempt has been made to identify the factors responsible for this persistent trend and to quantify these factors. This paper uses the Uganda Demographic and Health Surveys (UDHS) of 2006 and 2011, to examine the contribution contraceptive use, marriage and postpartum infecundability on one hand and Total Fertility Rate (TFR) on the other.We constructed a database using the Woman’s Questionnaire from the UDHS 2006 and 2011. We then apply Bongaarts aggregate fertility model procedures to derive estimates of total fertility rate for the different socioeconomic groups.The findings indicate that a woman’s contraceptive behavior; marriage status and postpartum infecundability , are important predictors of fertility outcomes. The results also show that higher education levels and urban residence are consistently associated with lower fertility rates and are positively associated with contraceptive use. Other key predictors of fertility include: wealth status, and region of residence.The country needs to scale-up target interventions that are aimed at uplifting the education status of women and improving their economic wellbeing, because such interventions have a positive impact on fertility reduction and on improving maternal and reproductive health outcomes. Reduction of total fertility rates (TFR) is a key determinant of overall reduction in population growth and transition from high to low fertility. These in turn may have important consequences for economic growth, poverty reduction, and improved health and nutrition outcomes . Uganda Socio-demographic characteristics of mothers like poor educational status, absence of income, rural place of birth, early marriage, and other variables like history of child death, negative husbands’ attitude towards contraceptive use, poor educational status of husbands, need for additional children, were found to have significant association with high fertility in Ethiopia –8.Other than contraceptive use, the fertility differences between population groups can be explained by the variations in other proximate determinants; notably exposure to the risk of pregnancy, and abstinence after delivery . Unmet nThere is now widespread agreement on the importance of men’s role in reproductive decision-making. Several studies have argued that fertility preferences and their translation into behavior differ between polygamous and monogamous unions . StudiesYoung women aspire to have an ideal ordering of events that places finishing education before getting married and having children, but this is often not easily attained , 15. TheWhile recent studies have indicated that fertility has remained high in Uganda, no systematic attempt has been made to identify the factors responsible for this persistent trend and to quantify these factors. Therefore the current study was based on the Bongaarts aggregate fertility model using data from two most recent DHS surveys of the Uganda: 2006 and 2011. This study explored the key drivers of fertility differences, including systematic analysis of the key proximate determinants, namely: contraception, marriage and postpartum insusceptibility.As noted earlier the data were extracted from the Uganda Demographic and Health Surveys conducted in 2006 and 2011. Approval for UDHS data utilized for this study was obtained from the data originator, ICF Macro International U.S.A before the data was extracted from their web platform. At the point of data collection by the data originators, an informed consent was sought from all the study participants after detailed description of all the issues related to the study were passed across to the respondents. Eligible respondents who did not want to participate in the study were excluded from the survey. Each consenting participants was made to sign appropriate agreement form before the commencement of the interview. These surveys employed nationally representative samples, which were based on a two-stage stratified sample of households.These types of surveys generally provide information on basic national indicators of social development. The present study utilised these data in order to fit the aggregate fertility model, thereby assessing the contribution of contraception, marriage and postpartum infecundability to fertility in Uganda. The fertility estimates were also disaggregated by a number of selected variables, namely: education, religion, residence, region of residence, and wealth index.A total of 8674 and 8531 women of ages 15–49 from the 2006 and 2011 Uganda Demographic and Health Surveys respectively were selected for the current study. In 2011 the household response rate was 97.4 % while the eligible women response rate was 93.8 %. Furthermore in 2006 the household response rate was 97.5 % while the eligible women response rate was 94.7 %. These samples are considered adequate to enable analyses and comparisons that would be useful in the identification of socio-economic and regional foci that could guide fertility and population policy interventions in Uganda.The Bongaarts model is adopted to quantify the contribution of the proximate determinants to fertility. Bongaarts’ original model included four proximate determinants: marriage, postpartum infecundability, abortion and contraception. In a later paper, Bongaarts added a fifth determinant, pathological sterility , 19. Them is the index of proportion married, Ci is the index of lactational infecundability, Ca is the index of abortion, Cc is the index of contraception and TF is total fecundity. The indices can only take values between 0 and 1. When there is no fertility-inhibiting effect of a given intermediate fertility variable, the corresponding index equals 1. If the fertility inhibition is complete, the index equals 0. These indices can be estimated from measures of the proximate variables. Although this aggregate version of the model is the most widely used, there is also an age-specific version that calculates the effects separately for each five year age group from 15–19 to 45–49 [where Cto 45–49 .The age-specific proportions of determine the index of marriage currently married among females. The index Cm is not simply equal to the proportion of all women of reproductive age that is married because the fertility impact of marriage also depends on the age distribution of married women. Married women in the central childbearing years contribute more to the TFR than the youngest or oldest women because the age-specific marital fertility rates reach their maximum in the central childbearing ages. The index is equal to the ratio of the total fertility rate to the total marital fertility rate. The index of marriage equals one when all women of reproductive age are in a union and zero when no women are in a union. Marriage in this context refers to both formal marriage and consensual unions. Implicit in the use of the index is the assumption that only women in union are exposed to the risk of childbirth. This assumption does not hold reasonably well in Uganda where childbearing outside of marriage do exist.Where: m(a) = age-specific proportions of women currently married (it is got by dividing the number of married women of a particular age group by the number of women in the same age group); g(a) = age-specific marital fertility rate (is got by dividing the births of a particular age group (from married women) by the number of married women in the same age group).c) varies inversely with prevalence and use-effectiveness of contraception practiced by couples in the reproductive age groups. It incorporates both prevalence of contraceptive use and estimated effectiveness of the mix of methods used. It equals one if no form of contraception is used and zero if all fecund women use modern methods  that have a higher effectiveness [The index of contraception .The index of postpartum infecundability is a measure of the inhibiting effect of breastfeeding or abstinence on fertility in the population . The indWhere: i = the mean duration of postpartum infecundability measured in months. According to Bongaarts, without lactation, a typical average birth interval is estimated at 20 months, and with lactation it equals the average total duration of the infecund period plus 18.5 months.Abortion is illegal in Uganda. The total abortion rate can be used to assess the relationship between induced abortions and fertility. This rate is equivalent to the TFR but includes only induced abortions (rather than births) in the numerator. Because of lack of reliable data for induced abortion, the index of abortion was estimated to 1. The difficulty of getting such data was reported by Bongaarts , and accThe index of abortion is estimated using the formula below;Where u = Prevalence contraceptive use; b = Average number of births averted per induced abortion and b = 0.4 (1 + u); b = 0.4 when u = 0 and b = 0.8 when u = 1.0.TA = Total abortion (Average number of induced abortions per woman at the end of the reproductive period if induced abortion rates remains at prevailing levels throughout the reproductive period). Therefore, C a = 1.0 if the TA is 0. Therefore the Total Abortion rate in this study is assumed to be 1.0.When all indices equal one, fertility is at its biological maximum. Based on studies of historical populations with the highest recorded fertility, Bongaarts recommends using 15.3 as the maximum number of births per woman; this is referred to as the total fecundity rate . This vaTable The distribution of the study population by marital status shows that the proportion currently married was 62.6 % in 2006 and this stayed nearly the same in 2011, while those who were single at the time of the survey were only about 24 % in the two surveys. About one quarter of the women respondents in both 2006 and also in 2011 were in the richest wealth quintile, while about 42 % of the respondents were in the lowest two wealth quintiles in both years. Similarly those women in the middle wealth quintile held steadily at about 19 % in both 2006 and 2011. The distribution of the study population by age distribution clearly reflects a similar distribution for both surveys. About one fifth of the study population was aged 15–19 years and the proportions at each age group gradually declined reaching about 7 % in the age group 45–49 years.Table Tables Results presented in Tables The rural/urban residential distribution shows that in 2006 and 2011, the contraceptive use index was better for urban areas relative to rural areas, with rural areas exhibiting some slight improvement compared to the urban areas at the two survey periods. The findings also suggest that contraceptive use was slightly higher among Protestants and Moslem religions in 2006 (Cc = 0.78 and 0.71), respectively. In 2011 there were some improvements in contraceptive use and some differences with Protestants and Other religions depicting better contraceptive use indicators (Cc = 0.68 and 0.66), respectively. In 2011, Moslems had a Cc of 0.71, which was the same as that of 2006.Contraceptive index (Cc) was least among women in the richest wealth quintile for both 2006 and 2011 (Cc = 0.54), for both years respectively. Women in the poorest wealth quintile had the highest Cc index of 0.85 and 0.89 in 2006 and 2011, respectively, suggesting that contraceptive use effect on fertility was least among the poorest categories of the population compared to the richest individuals. Finally, the contribution of contraceptives to fertility inhibition was highest among women with a secondary and higher education compared to other education categories in both 2006 and 2011. The value of Cc held steadily at 0.56 at both survey periods, while slight improvements were observed among those with no education and those with a primary education during the same period.The index of marriage had the greatest inhibiting effect on fertility in Uganda in 2006 (Cm = 0.72). As is the case with the index of contraception, a higher value of Cm index is consistent low effective impact on fertility reduction. The value of Cm was the same in 2011 and had nearly same effect as Cc in the same year. The high contribution of the index of marriage to fertility reduction was especially substantial in Kampala region (Cm = 0.57), compared to other regions of the country. Eastern region depicted the highest index on marriage at 0.76. Similarly the index of marriage (Cm) was least among those residing in the rural areas (Cm = 0.78) compared to those in urban areas (Cm = 0.59).Concerning the wealth index, the index of marriage was least among women in the richest wealth quintile (Cm = 0.58) in both 2006 and in 2011. The marriage index was highest among those in the poorest category at 0.78 and 0.82 in 2006 and 2011, respectively. Overall the findings in Tables The index of postpartum infecundability provided the least overall inhibiting effect on fertility compared to Cm and Cc in 2011 (0.76), but performed better in 2006 (Ci = 0.74). The results generally show that religious variations in the value of this index were minimal, ranging from 0.72 among Catholic’s to 0.76 among Moslems in 2006, and from 0.75 to 0.77 among the same groups, respectively in 2011.Regional differences in the Ci suggest that Kampala, the capital city, had the highest value of the index at 0.81 and 0.83 in 2006 and 2011, respectively, while Northern region had the least value of the index at 0.79 and 0.72 at the two respective periods. The rural urban dimensions show that urban areas consistently had a high value of Ci compared to rural areas. In 2006 the Ci for urban areas of Uganda was 0.79 and was estimated at 0.81 in 2011.The differences in the value of Ci were apparent for the various wealth index categories, but clearly, there was a direct relationship between wealth quintile and the value of Ci for both 2006 and 2011. This direct relationship was also observed for the education level attainment categories. Women with a secondary and higher education depicted the highest values of Ci (0.82 in 2011 and 0.80 in 2006) compared to those with no education (0.72 in 2011 and 0.71 in 2006).The aggregate fertility model findings presented in Tables Regional differences in TFR were observed for both 2006 and 2011. Kampala district had the least TFR estimate of 3.5 in 2006, increasing to 3.9 in 2011, while Northern region had the highest TFR of 7.4 in 2006 declining to 6.8 in 2011. The rural/urban patterns show constant fertility at 6.9 children in rural areas for both 2006 and 2011. With regard to wealth index, the results in this paper suggest that women in the poorest wealth quintile had the highest TFR while those within the richest wealth quintiles had the least fertility outcomes. This latter finding is consistent for both 2006 and 2011. Similarly, women with a secondary and higher education had the least TFR compared to those with no education or those with a primary level of education.This study sought to establish the contribution of contraception, marriage and postpartum infecundability to the total fertility rate in Uganda. Using the Aggregate Fertility Model framework , 23, it Furthermore the index of abortion was set at 1.0 in the model due to lack of data on this indicator. It is therefore important that future data collection undertakings should include questions that can allow for investigation using this fourth parameter. Nonetheless it was found in the current study that background factors including: region, residence, education, religion, and wealth status also have an influence on fertility outcomes, given that they may influence the levels of the proximate determinants.The major limitation of this study relates to the secondary nature of the data that were used. Invariably many events captured through a retrospective inquiry are often susceptible to recall bias and memory lapse. One major limitation of this study is the assumption that the index of abortion is unity and has no substantial effect on fertility estimates. This was because these data on abortion were missing and were not available for any comparable population. We observe, however that in societies that are predominantly traditional rural in nature, the assumption that induced abortion plays a minimal role would still be appropriate, and this is what guided this study. Moreover, according Bongaarts–the author of the method use , modelinTherefore, inclusion of background factors in future studies has the potential and advantage of informing policy and programming on the segments of the population that require policy and programmatic intervention. The country needs to scale-up target interventions that are aimed at uplifting the education status of women and improving their economic wellbeing, because such interventions have a positive impact on fertility outcomes. In this regard, population groups with poor indices of proximate determinants of fertility would be of particular interest.