The automation of protein crystal presentation for x ray diffraction experiments using standing acoustic waves in a microfluidic chip environment

As the pressure continues to grow on Diamond and the world's synchrotrons for higher throughput of diffraction experiments, new and novel techniques are required for presenting micron dimension crystals to the X ray beam. Currently this task is both labour intensive and primarily a serial process. Diffraction measurements typically take milliseconds but sample preparation and presentation can reduce throughput down to 4 measurements an hour. With beamline waiting times as long as two years it is of key importance for researchers to capitalize on available beam time, generating as much data as possible. Other approaches detailed in the literature [1] [2] [3] are very much skewed towards automating, with robotics, the actions of a human protocols. The work detailed here is the development and discussion of a bottom up approach relying on SSAW self assembly, including material selection, microfluidic integration and tuning of the acoustic cavity to order the protein crystals.

The automation of protein crystal presentation for X ray diffraction experiments using standing acoustic waves in a microfluidic chip environment

As the pressure continues to grow on Diamond and the world's synchrotrons for higher throughput of diffraction experiments, new and novel techniques are required for presenting micron dimension crystals to the X ray beam. Currently this task is both labour intensive and primarily a serial process. Diffraction measurements typically take milliseconds but sample preparation and presentation can reduce throughput down to 4 measurements an hour. With beamline waiting times as long as two years it is of key importance for researchers to capitalize on available beam time, generating as much data as possible. Other approaches detailed in the literature [1] [2] [3] are very much skewed towards automating, with robotics, the actions of a human protocols. The work detailed here is the development and discussion of a bottom up approach relying on SSAW self assembly, including material selection, microfluidic integration and tuning of the acoustic cavity to order the protein crystals.

Receptors, sparks and waves in a fire-diffuse-fire framework for calcium release

Calcium ions are an important second messenger in living cells. Indeed calcium signals in the form of waves have been the subject of much recent experimental interest. It is now well established that these waves are composed of elementary stochastic release events (calcium puffs or sparks) from spatially localised calcium stores. The aim of this paper is to analyse how the stochastic nature of individual receptors within these stores combines to create stochastic behaviour on long timescales that may ultimately lead to waves of activity in a spatially extended cell model. Techniques from asymptotic analysis and stochastic phase-plane analysis are used to show that a large cluster of receptor channels leads to a release probability with a sigmoidal dependence on calcium density. This release probability is incorporated into a computationally inexpensive model of calcium release based upon a stochastic generalization of the Fire-Diffuse-Fire (FDF) threshold model. Numerical simulations of the model in one and two dimensions (with stores arranged on both regular and disordered lattices) illustrate that stochastic calcium release leads to the spontaneous production of calcium sparks that may merge to form saltatory waves. Illustrations of spreading circular waves, spirals and more irregular waves are presented. Furthermore, receptor noise is shown to generate a form of array enhanced coherence resonance whereby all calcium stores release periodically and simultaneously.

Ocean waves across the Arctic: attenuation due to dissipation dominates over scattering for periods longer than 19 s

The poorly understood attenuation of surface waves in sea ice is generally attributed to the combination of scattering and dissipation. Scattering and dissipation have very different effects on the directional and temporal distribution of wave energy, making it possible to better understand their relative importance by analysis of swell directional spreading and arrival times. Here we compare results of a spectral wave model – using adjustable scattering and dissipation attenuation formulations – with wave measurements far inside the ice pack. In this case, scattering plays a negligible role in the attenuation of long swells. Specifically, scattering-dominated attenuation would produce directional wave spectra much broader than the ones recorded, and swell events arriving later and lasting much longer than observed. Details of the dissipation process remain uncertain. Average dissipation rates are consistent with creep effects but are 12 times those expected for a laminar boundary layer under a smooth solid ice plate.

Some misspecfication problems in long-memory time series models

Doctor of Philosophy in Mathematics

TECTONIC CONTROLS ON FLUVIAL LANDSCAPE DEVELOPMENT IN CENTRAL-EASTERN PORTUGAL: INSIGHTS FROM LONG PROFILE TRIBUTARY STREAM ANALYSES

This study examines the long profiles of tributaries of the Tejo (Tagus) and Zêzere rivers in central eastern Portugal (West Iberia) in order to provide new insights into the patterns, timing and controls on drainage development during the Pleistocene to Holocene incision stage. The long profiles were extracted from lower order tributary streams associated with the trunk drainage of the Tejo River and one main tributary, the Zêzere River (Fig. 1). These streams flow through a landscape strongly influenced by variations in bedrock lithology (mainly granites and metasediments), fault structures delimiting crustal blocks with distinct uplift rates, and a base-level lowering history (tectonic uplift / eustatic). The long profiles of the tributaries of the Tejo and Zêzere rivers record a series of transient and permanent knickpoints. The permanent knickpoints have direct correlation with the bedrock strength, corresponding to the outcropping of very hard quartzites or to the transition from softer (slates/metagreywaques) to harder (granite) basement. The analyzed streams/rivers record also an older transient knickpoint/knickzone separating: a) an upstream relict graded profile, with lower steepness and higher concavity, that reflects a long period of quasi-equilibrium conditions reached after the beginning of the incision stage; and b) a downstream reach displaying a rejuvenated long profile, with steeper gradient and lower concavity, particularly for the final segment, which is often convex (Fig. 2). The rejuvenated reaches testify the upstream propagation of several incision waves that are the response of each stream to continuous or increasing crustal uplift and dominant periods of base-level lowering by the trunk drainages, coeval of low sea level conditions. The long profiles and their morphological configurations enabled spatial and relative temporal patterns of incision to be quantified for each individual tributary stream. The incision values of streams flowing in uplifted blocks of the Portuguese Central Range (PCR) (ca.380-280 m) indicate differential uplift and are higher than the incision values of streams flowing on the adjacent South Portugal planation surface – the Meseta (ca. 200 m). The normalized steepness index, calculated using the method of Wobus et al. (2006), proved to be sensitive to active tectonics, as lower ksn values were found in relict graded profiles of streams located in less uplifted blocks, (e.g. Sertã stream in the PCR), or in those flowing through tectonic depressions. Fig. 1 – Geological map of the study area. 1 – fluvial terraces (Pleistocene); 2 – sedimentary cover (Paleogene and Neogene); 3 – slates and metasandstones (Devonian); 4 – slates and quartzites (Silurian); 5 – quartzites (Ordovician); 6 – slates and metagreywackes (Precambrian to Cambrian); 7 – slates, metagreywackes and limestones (Precambrian); 8 – granites and ortogneisses; 9 – diorites and gabros; 10 - fault. SFf – Sobreira Formosa fault; Sf – Sertã fault; Pf – Ponsul fault; Gf – Grade fault. The differential uplift indicated by the distribution of the ksn values and by the fluvial incision was likely accumulated on a few major faults, as the Sobreira Formosa fault (SFf), thus corroborating the tectonic activity of these faults. Due to the fact that the relict graded profiles can be correlated with other geomorphic references documented in the study area, namely the T1 terrace of the Tagus River (with an age of ca. 1 Myr), the following incision rates can be estimated: a) for the studied streams located in uplifted blocks of the PCR, 0.38 m/kyr to 0.28 m/kyr; b) for the streams flowing on the South Portugal planation surface, 0.20 m/kyr. The differential uplift inferred between crustal blocks in the study area corroborates the neotectonic activity of the bordering faults, which has been proposed in previous studies based upon less robust data. Fig. 2 – Longitudinal profile of the Nisa stream a tributary of the Tejo River. Note the equilibrium relict profile upstream the older transient knickpoint (hatched line) and the downstream rejuvenated profile (continuous line). Legend: tKP – transient knickpoint; rKp – resistant knickpoint; Mt – schist and phyllite; Gr – granite; Hf – hornfels; Og – orthogneisse. In the inset Distance – Slope plots, fill circles correspond to the relict graded profile, crosses correspond to the rejuvenated profile located downstream the older transient knickpoint (tKP).

Tectonic and lithological controls on fluvial landscape development in central-eastern Portugal: Insights from long profile tributary stream analyses

This study examines the long profiles of tributaries of the Tagus and Zêzere rivers in Portugal (West Iberia) in order to provide new insights into patterns, timing, and controls on drainage development during the Quaternary incision stage. The studied streams are incised into a relict culminant fluvial surface, abandoned at the beginning of the incision stage. The streams flow through a landscape with bedrock variations in lithology (mainly granites and metasediments) and faulted blocks with distinct uplift rates. The long profiles of the analyzed streams record an older transitory knickpoint/knickzone separating (1) an upstream relict graded profile, with lower steepness and higher concavity, that reflects a long period of quasi-equilibrium conditions reached after the beginning of the incision stage, and (2) a downstream rejuvenated long profile, with steeper gradient and lower concavity, particularly for the final reach, which is often convex. The rejuvenated reaches testify to the upstream propagation of several incision waves, interpreted as the response of each stream to increasing crustal uplift and prolonged periods of base-level lowering by the trunk drainages, coeval with low sea level conditions. The morphological configurations of the long profiles enabled spatial and relative temporal patterns of incisions to be quantified. The incision values of streams flowing on the Portuguese Central Range (PCR; ca. 380–150 m) are variable but generally higher than the incision values of streams flowing on the adjacent South Portugal Planation Surface (SPPS; ca. 220–110 m), corroborating differential uplift of the PCR relative to the SPPS. Owing to the fact that the relict graded profiles can be correlated with the Tagus River T1 terrace (1.1–0.9 My) present in the study area, incision rates can be estimated (1) for the streams located in the PCR, 0.38–0.15 m/ky and (2) for the streams flowing on the SPPS, 0.22–0.12 m/ky. The differential uplift inferred in the study area supports the neotectonic activity of the bordering faults, as proposed in previous studies based upon other geological evidence.

Tectonic and lithological controls on fluvial landscape development in central-eastern Portugal: Insights from long profile tributary stream analyses

This study examines the long profiles of tributaries of the Tagus and Zêzere rivers in Portugal (West Iberia) in order to provide new insights into patterns, timing, and controls on drainage development during the Quaternary incision stage. The studied streams are incised into a relict culminant fluvial surface, abandoned at the beginning of the incision stage. The streams flow through a landscape with bedrock variations in lithology (mainly granites and metasediments) and faulted blocks with distinct uplift rates. The long profiles of the analyzed streams record an older transitory knickpoint/knickzone separating (1) an upstream relict graded profile, with lower steepness and higher concavity, that reflects a long period of quasi-equilibrium conditions reached after the beginning of the incision stage, and (2) a downstream rejuvenated long profile, with steeper gradient and lower concavity, particularly for the final reach, which is often convex. The rejuvenated reaches testify to the upstream propagation of several incision waves, interpreted as the response of each stream to increasing crustal uplift and prolonged periods of base-level lowering by the trunk drainages, coeval with low sea level conditions. The morphological configurations of the long profiles enabled spatial and relative temporal patterns of incisions to be quantified. The incision values of streams flowing on the Portuguese Central Range (PCR; ca. 380–150 m) are variable but generally higher than the incision values of streams flowing on the adjacent South Portugal Planation Surface (SPPS; ca. 220–110 m), corroborating differential uplift of the PCR relative to the SPPS. Owing to the fact that the relict graded profiles can be correlated with the Tagus River T1 terrace (1.1–0.9 My) present in the study area, incision rates can be estimated (1) for the streams located in the PCR, 0.38–0.15 m/ky and (2) for the streams flowing on the SPPS, 0.22–0.12 m/ky. The differential uplift inferred in the study area supports the neotectonic activity of the bordering faults, as proposed in previous studies based upon other geological evidence.

Resonant metamaterials for the control of Rayleigh waves

Since their emergence, locally resonant metamaterials have found several applications for the control of surface waves, from micrometer-sized electronic devices to meter-sized seismic barriers. The interaction between Rayleigh-type surface waves and resonant metamaterials has been investigated through the realization of locally resonant metasurfaces, thin elastic interfaces constituted by a cluster of resonant inclusions or oscillators embedded near the surface of an elastic waveguide. When such resonant metasurfaces are embedded in an elastic homogeneous half-space, they can filter out the propagation of Rayleigh waves, creating low-frequency bandgaps at selected frequencies. In the civil engineering context, heavy resonating masses are needed to extend the bandgap frequency width of locally resonant devices, a requirement that limits their practical implementations. In this dissertation, the wave attenuation capabilities of locally resonant metasurfaces have been enriched by proposing (i) tunable metasurfaces to open large frequency bandgaps with small effective inertia, and by developing (ii) an analytical framework aimed at studying the propagation of Rayleigh waves propagation in deep resonant waveguides. In more detail, inertial amplified resonators are exploited to design advanced metasurfaces with a prescribed static and a tunable dynamic response. The modular design of the tunable metasurfaces allows to shift and enlarge low-frequency spectral bandgaps without modifying the total inertia of the metasurface. Besides, an original dispersion law is derived to study the dispersive properties of Rayleigh waves propagating in thick resonant layers made of sub-wavelength resonators. Accordingly, a deep resonant wave barrier of mechanical resonators embedded inside the soil is designed to impede the propagation of seismic surface waves. Numerical models are developed to confirm the analytical dispersion predictions of the tunable metasurface and resonant layer. Finally, a medium-size scale resonant wave barrier is designed according to the soil stratigraphy of a real geophysical scenario to attenuate ground-borne vibration.

Mechanisms Predisposing Penile Fracture And Long-term Outcomes On Erectile And Voiding Functions.

Purpose. To determine the mechanisms predisposing penile fracture as well as the rate of long-term penile deformity and erectile and voiding functions. Methods. All fractures were repaired on an emergency basis via subcoronal incision and absorbable suture with simultaneous repair of eventual urethral lesion. Patients' status before fracture and voiding and erectile functions at long term were assessed by periodic follow-up and phone call. Detailed history included cause, symptoms, and single-question self-report of erectile and voiding functions. Results. Among the 44 suspicious cases, 42 (95.4%) were confirmed, mean age was 34.5 years (range: 18-60), mean follow-up 59.3 months (range 9-155). Half presented the classical triad of audible crack, detumescence, and pain. Heterosexual intercourse was the most common cause (28 patients, 66.7%), followed by penile manipulation (6 patients, 14.3%), and homosexual intercourse (4 patients, 9.5%). Woman on top was the most common heterosexual position (n = 14, 50%), followed by doggy style (n = 8, 28.6%). Four patients (9.5%) maintained the cause unclear. Six (14.3%) patients had urethral injury and two (4.8%) had erectile dysfunction, treated by penile prosthesis and PDE-5i. No patient showed urethral fistula, voiding deterioration, penile nodule/curve or pain. Conclusions. Woman on top was the potentially riskiest sexual position (50%). Immediate surgical treatment warrants long-term very low morbidity.

Up-regulation Of Dnam-1 And Nkp30, Associated With Improvement Of Nk Cells Activation After Long-term Culture Of Mononuclear Cells From Patients With Ovarian Neoplasia.

This study aimed at evaluating the functional activation and activating receptors expression on resting, short- and long-term NK and NK-like T cells from blood of ovarian neoplasia patients. Blood from patients with adnexal benign alterations (n = 10) and ovarian cancer (grade I-IV n = 14) were collected after signed consent. Effector cells activation was evaluated by the expression of the CD107a molecule. Short-term culture was conducted overnight with IL-2 and long-term culture for 21 days, by a method designed to expand CD56(+) lymphocytes. Short-term culture significantly increased NK cells activation compared to resting NK cells (p<0.05), however, the long-term procedure supported an even higher increase (p<0.001). Resting NK-like T cells showed poor activation, which was not altered by the culture procedures. The long-term culture effectively increased the expression of the activating receptors on NK and NK-like T cells, either by increasing the number of cells expressing a given receptor and/or by up-regulating their expression intensity. As a conclusion, the long-term culture system employed, resulted in a high number of functional NK cells. The culture system was particularly efficient on the up-regulation of NKp30 and DNAM-1 receptors on NK cells.

Genetic Predictors Of Long-term Response To Growth Hormone (gh) Therapy In Children With Gh Deficiency And Turner Syndrome: The Influence Of A Socs2 Polymorphism.

There is great interindividual variability in the response to GH therapy. Ascertaining genetic factors can improve the accuracy of growth response predictions. Suppressor of cytokine signaling (SOCS)-2 is an intracellular negative regulator of GH receptor (GHR) signaling. The objective of the study was to assess the influence of a SOCS2 polymorphism (rs3782415) and its interactive effect with GHR exon 3 and -202 A/C IGFBP3 (rs2854744) polymorphisms on adult height of patients treated with recombinant human GH (rhGH). Genotypes were correlated with adult height data of 65 Turner syndrome (TS) and 47 GH deficiency (GHD) patients treated with rhGH, by multiple linear regressions. Generalized multifactor dimensionality reduction was used to evaluate gene-gene interactions. Baseline clinical data were indistinguishable among patients with different genotypes. Adult height SD scores of patients with at least one SOCS2 single-nucleotide polymorphism rs3782415-C were 0.7 higher than those homozygous for the T allele (P < .001). SOCS2 (P = .003), GHR-exon 3 (P= .016) and -202 A/C IGFBP3 (P = .013) polymorphisms, together with clinical factors accounted for 58% of the variability in adult height and 82% of the total height SD score gain. Patients harboring any two negative genotypes in these three different loci (homozygosity for SOCS2 T allele; the GHR exon 3 full-length allele and/or the -202C-IGFBP3 allele) were more likely to achieve an adult height at the lower quartile (odds ratio of 13.3; 95% confidence interval of 3.2-54.2, P = .0001). The SOCS2 polymorphism (rs3782415) has an influence on the adult height of children with TS and GHD after long-term rhGH therapy. Polymorphisms located in GHR, IGFBP3, and SOCS2 loci have an influence on the growth outcomes of TS and GHD patients treated with rhGH. The use of these genetic markers could identify among rhGH-treated patients those who are genetically predisposed to have less favorable outcomes.

Long-term Postoperative Atrophy Of Contralateral Hippocampus And Cognitive Function In Unilateral Refractory Mtle With Unilateral Hippocampal Sclerosis.

This study aimed to evaluate long-term atrophy in contralateral hippocampal volume after surgery for unilateral MTLE, as well as the cognitive outcome for patients submitted to either selective transsylvian amygdalohippocampectomy (SelAH) or anterior temporal lobe resection (ATL). We performed a longitudinal study of 47 patients with MRI signs of unilateral hippocampal sclerosis (23 patients with right-sided hippocampal sclerosis) who underwent surgical treatment for MTLE. They underwent preoperative/postoperative high-resolution MRI as well as neuropsychological assessment for memory and estimated IQ. To investigate possible changes in the contralateral hippocampus of patients, we included 28 controls who underwent two MRIs at long-term intervals. The volumetry using preoperative MRI showed significant hippocampal atrophy ipsilateral to the side of surgery when compared with controls (p<0.0001) but no differences in contralateral hippocampal volumes. The mean postoperative follow-up was 8.7 years (± 2.5 SD; median=8.0). Our patients were classified as Engel I (80%), Engel II (18.2%), and Engel III (1.8%). We observed a small but significant reduction in the contralateral hippocampus of patients but no volume changes in controls. Most of the patients presented small declines in both estimated IQ and memory, which were more pronounced in patients with left TLE and in those with persistent seizures. Different surgical approaches did not impose differences in seizure control or in cognitive outcome. We observed small declines in cognitive scores with most of these patients, which were worse in patients with left-sided resection and in those who continued to suffer from postoperative seizures. We also demonstrated that manual volumetry can reveal a reduction in volume in the contralateral hippocampus, although this change was mild and could not be detected by visual analysis. These new findings suggest that dynamic processes continue to act after the removal of the hippocampus, and further studies with larger groups may help in understanding the underlying mechanisms.

Reasons For Brazilian Women To Switch From Different Contraceptives To Long-acting Reversible Contraceptives.

Long-acting reversible contraceptives (LARCs) include the copper-releasing intrauterine device (IUD), the levonorgestrel-releasing intrauterine system (LNG-IUS) and implants. Despite the high contraceptive efficacy of LARCs, their prevalence of use remains low in many countries. The objective of this study was to assess the main reasons for switching from contraceptive methods requiring daily or monthly compliance to LARC methods within a Brazilian cohort. Women of 18-50 years of age using different contraceptives and wishing to switch to a LARC method answered a questionnaire regarding their motivations for switching from their current contraceptive. Continuation rates were evaluated 1 year after method initiation. Sample size was calculated at 1040 women. Clinical performance was evaluated by life table analysis. The cutoff date for analysis was May 23, 2013. Overall, 1167 women were interviewed; however, after 1 year of use, the medical records of only 1154 women were available for review. The main personal reason for switching, as reported by the women, was fear of becoming pregnant while the main medical reasons were nausea and vomiting and unscheduled bleeding. No pregnancies occurred during LARC use, and the main reasons for discontinuation were expulsion (in the case of the IUD and LNG-IUS) and a decision to undergo surgical sterilization (in the case of the etonogestrel-releasing implant). Continuation rate was ~95.0/100 women/year for the three methods. Most women chose a LARC method for its safety and for practical reasons, and after 1 year of use, most women continued with the method.

Estimated Disability-adjusted Life Years Averted By Long-term Provision Of Long Acting Contraceptive Methods In A Brazilian Clinic.

What is the contribution of the provision, at no cost for users, of long acting reversible contraceptive methods (LARC; copper intrauterine device [IUD], the levonorgestrel-releasing intrauterine system [LNG-IUS], contraceptive implants and depot-medroxyprogesterone [DMPA] injection) towards the disability-adjusted life years (DALY) averted through a Brazilian university-based clinic established over 30 years ago. Over the last 10 years of evaluation, provision of LARC methods and DMPA by the clinic are estimated to have contributed to DALY averted by between 37 and 60 maternal deaths, 315-424 child mortalities, 634-853 combined maternal morbidity and mortality and child mortality, and 1056-1412 unsafe abortions averted. LARC methods are associated with a high contraceptive effectiveness when compared with contraceptive methods which need frequent attention; perhaps because LARC methods are independent of individual or couple compliance. However, in general previous studies have evaluated contraceptive methods during clinical studies over a short period of time, or not more than 10 years. Furthermore, information regarding the estimation of the DALY averted is scarce. We reviewed 50 004 medical charts from women who consulted for the first time looking for a contraceptive method over the period from 2 January 1980 through 31 December 2012. Women who consulted at the Department of Obstetrics and Gynaecology, University of Campinas, Brazil were new users and users switching contraceptive, including the copper IUD (n = 13 826), the LNG-IUS (n = 1525), implants (n = 277) and DMPA (n = 9387). Estimation of the DALY averted included maternal morbidity and mortality, child mortality and unsafe abortions averted. We obtained 29 416 contraceptive segments of use including 25 009 contraceptive segments of use from 20 821 new users or switchers to any LARC method or DMPA with at least 1 year of follow-up. The mean (± SD) age of the women at first consultation ranged from 25.3 ± 5.7 (range 12-47) years in the 1980s, to 31.9 ± 7.4 (range 16-50) years in 2010-2011. The most common contraceptive chosen at the first consultation was copper IUD (48.3, 74.5 and 64.7% in the 1980s, 1990s and 2000s, respectively). For an evaluation over 20 years, the cumulative pregnancy rates (SEM) were 0.4 (0.2), 2.8 (2.1), 4.0 (0.4) and 1.3 (0.4) for the LNG-IUS, the implants, copper IUD and DMPA, respectively and cumulative continuation rates (SEM) were 15.1 (3.7), 3.9 (1.4), 14.1 (0.6) and 7.3 (1.7) for the LNG-IUS, implants, copper IUD and DMPA, respectively (P < 0.001). Over the last 10 years of evaluation, the estimation of the contribution of the clinic through the provision of LARC methods and DMPA to DALY averted was 37-60 maternal deaths; between 315 and 424 child mortalities; combined maternal morbidity and mortality and child mortality of between 634 and 853, and 1056-1412 unsafe abortions averted. The main limitations are the number of women who never returned to the clinic (overall 14% among the four methods under evaluation); consequently the pregnancy rate could be different. Other limitations include the analysis of two kinds of copper IUD and two kinds of contraceptive implants as the same IUD or implant, and the low number of users of implants. In addition, the DALY calculation relies on a number of estimates, which may vary in different parts of the world. LARC methods and DMPA are highly effective and women who were well-counselled used these methods for a long time. The benefit of averting maternal morbidity and mortality, child mortality, and unsafe abortions is an example to health policy makers to implement more family planning programmes and to offer contraceptive methods, mainly LARC and DMPA, at no cost or at affordable cost for the underprivileged population. This study received partial financial support from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), grant # 2012/12810-4 and from the National Research Council (CNPq), grant #573747/2008-3. B.F.B., M.P.G., and V.M.C. were fellows from the scientific initiation programme from FAPESP. Since the year 2001, all the TCu380A IUD were donated by Injeflex, São Paulo, Brazil, and from the year 2006 all the LNG-IUS were donated by the International Contraceptive Access Foundation (ICA), Turku, Finland. Both donations are as unrestricted grants. The authors declare that there are no conflicts of interest associated with this study.