Serum transitional levels of IL-6 (p < 0 001), sIL-2R (p <

Serum transitional levels of IL-6 (p < 0.001), sIL-2R (p < 0.001), sTNFRII (p < 0.001), and G-CSF (p < 0.001) reflect disease severity. In Non-responders, average levels of IL-6 at day 7 (43.5 vs. 13.8 pg/ml, p < 0.001) and average levels of BYL719 inhibitor sIL-2R at day 14 (21.3 vs. 3.31 pg/ml, p = 0.014) were significantly higher than those in Responders.\n\nConclusion: CsA treatment effectively reduced the persisting serum inflammatory cytokines in most of the IVIG-resistant KD patients. Soluble IL-2R suppression

implies a mechanism explaining the effects of CsA. (C) 2012 Elsevier Ltd. All rights reserved.”
“The family of superoxide dismutases (SODs) are well known for their antioxidant actions exerted by catalyzing the conversion of O(2)(-) into H(2)O(2) plus oxygen. The importance of this action is revealed by the multiple phenotypic deficits exhibited by a variety of organisms that have been made to lack one or more of the SODs. Never the less there

have been reports of deleterious consequences caused by overproduction of SOD. Several explanations have been proposed for these counter intuitive effects; one of which is that elevated Sapitinib datasheet SOD causes increased formation of H(2)O(2). The reasons for dismissing this explanation are explored.”
“Objective: To investigate the effects of pioglitazone on ovarian stimulation, in vitro fertilization (IVF) outcome, and intraovarian stromal blood flow in patients with polycystic ovary syndrome (PCOS).\n\nDesign: Prospective, randomized, controlled trial.\n\nSetting: University-affiliated infertility clinic.\n\nPatient(s): Sixty infertile patients with PCOS resistant to clomiphene citrate.\n\nIntervention(s): BB-94 price An IVF protocol involving use of a gonadotropin-releasing hormone (GnRH) antagonist and oral contraceptive pretreatment. Patients were randomized into two groups: pioglitazone (30 mg daily) in the study group or placebo in the control group, commenced on the day on which oral contraceptive

intake began.\n\nMain Outcome Measure(s): Ovarian stimulation, IVF outcome, and resistance index of the intraovarian stromal artery.\n\nResult(s): The resistance index value on the day of human chorionic gonadotropin (hCG) injection in the study group was statistically significantly higher in the study group than in controls. The serum estradiol level and number of retrieved oocytes were statistically significantly lower in women treated with pioglitazone. However, the number of mature oocytes and fertilized oocytes, and the clinical pregnancy rate were similar in both groups.\n\nConclusion(s): Pioglitazone therapy reduced intraovarian stromal blood flow and might be beneficial in improving both the response to ovarian stimulation and IVF outcome in PCOS patients. (Fertil Steril (R) 2010; 94: 236-41. (C) 2010 by American Society for Reproductive Medicine.

We found thousands of transcripts with transposable element inser

We found thousands of transcripts with transposable element insertions in or near the transcript and that the presence of a transposable element in or near a transcript is significantly associated with reductions in expression. We estimate that within this example Prexasertib molecular weight population, similar to 2.2% of transcripts have a transposable element insertion, which significantly reduces expression in the line containing the transposable element. We also find that transcripts with insertions within 500 bp of the transcript show on average a

0.67 standard deviation decrease in expression level. These large decreases in expression level are most pronounced for transposable element insertions close to transcripts and the effect diminishes for more distant insertions. This work represents the first genome-wide analysis of gene expression variation due to transposable elements and suggests that transposable elements are an important class of mutation underlying expression variation in Drosophila and likely in other systems, given the ubiquity of these mobile elements in eukaryotic genomes.”
“Background: Several studies documented that lower scores On the Momingness-Eveningness Questionnaire (MEQ) are associated with a higher global seasonality of mood (GSS). As for the Modern Man artificial lighting predominantly extends HDAC activation evening

activity and exposure to light, and as evening bright light phase is selleck kinase inhibitor known to delay circadian rhythms, this chronic exposure could potentially lead to both lower Momingness as well as higher GSS. The aim of the study was to investigate if

the MEQ-GSS relationship holds in the Old Order Amish of Lancaster County, PA, a population that does not use network electrical light. Methods: 489 Old Order Amish adults (47.6% women), with average (SD) age of 49.7 (14.2) years, completed both the Seasonal Pattern Assessment Questionnaire (SPAQ) for the assessment of GSS, and MEQ. Associations between GSS scores and MEQ scores were analyzed using linear models, accounting for age, gender and relatedness by including the relationship matrix in the model as a random effect. Results: GSS was inversely associated with MEQ scores (p=0.006, adjusted). Limitations include a potential recall bias associated with self-report questionnaires and no actual light exposure measurements. Conclusion: We confirmed the previously reported inverse association between MEQ scores and lower seasonality of mood, for the first time in a population that does not use home network electrical lighting. This result suggests that the association is not a byproduct of exposure to network electric light, and calls for additional research to investigate mechanisms by which Momingness is negatively associated with seasonality. Published by Elsevier B.V.

and Phyla obtusa Audinet-Serville (both Coleoptera: Carabidae: Be

and Phyla obtusa Audinet-Serville (both Coleoptera: Carabidae: Bembidiini). We compared plant climbing behaviour, daily activity patterns, and trophic preferences between the two carabid species under laboratory conditions. Whereas no clear difference in trophic preference was observed, our results suggest temporal niche differentiation at the nychthemeron scale (a period of 24 consecutive hours), with one of the species being more diurnal and the other SN-38 more nocturnal, and spatial differentiation in their

habitat use at the plant stratum scale. Intra-specific variation suggests that micro-scale spatio-temporal niche differentiation could be mediated by behavioural plasticity in these two carabid species. We speculate that such behavioural plasticity may provide carabid beetles with a high adaptive potential in intensively managed agricultural areas.”
“Background: Medications with anticholinergic and sedative effects carry significant risks in older people. Adverse events arising from the use of these medications may also lead to hospitalization this website and contribute to length of stay. The Drug Burden Index (DBI) is a tool that measures a person’s total exposure to medications with anticholinergic and sedative properties, using the principles of dose response and maximal effect. Cumulative anticholinergic and sedative drug

burden measured using the DBI has been associated with clinically important outcomes in older people. The association between the DBI and hospitalization still remains relatively unknown.\n\nObjective: The main aim of this study was to evaluate the relationship between DBI and hospitalization in a population-based sample SRT2104 of community-dwelling older Finns over a 1-year period.\n\nMethods: The health status and medication use of 339 community-dwelling >= 75-year-old Finns were assessed in 2004. Data on hospitalizations over the following year were obtained from the national discharge register. Two different measures were used to assess hospitalizations in the study sample: (i) the proportion of hospitalized participants; and (ii) the number of hospital days per person-year. Estimates for the number

of hospital days per person-year and rate ratios (RRs) with 95% confidence intervals (CIs) were calculated using Poisson or negative binomial regression analysis.\n\nResults: A total of 127 participants (38%) were exposed to DBI medications; 27% had a low DBI (>0 to <1), and 11% had a high DBI (>= 1). The number of hospital days per person-year was 7.9 (95% CI 7.6, 8.3) for the unexposed participants (DBI = 0) and 13.4 (95% CI 12.8, 14.1) for the exposed participants (DBI >1); the age, gender and co-morbidity adjusted RR of hospital days per person-year between the exposed and unexposed participants was 1.26 (95% CI 1.18, 1.35). Between the low and high DBI groups, the difference in the number of hospital days per person-year was insignificant (p = 0.42).

P provide evidence for systematic harvesting and processing of e

P. provide evidence for systematic harvesting and processing of edible wild plants, including acorns and pine nuts. Analysis of oral pathology reveals this website an exceptionally high prevalence of caries (51.2% of teeth in adult dentitions), comparable to modern industrialized populations with a diet high in refined sugars and processed cereals. We infer that increased reliance on wild plants rich in fermentable carbohydrates and changes in food processing caused

an early shift toward a disease-associated oral microbiota in this population.”
“Atopic dermatitis (AD) is a common, chronic or chronically relapsing, severely pruritic and eczematous skin disease. AD is the second most frequently observed skin disease in dermatology clinics in Japan. Prevalence of childhood AD is 12-13% in mainland Japan; however, it is only half that (~6%) in children from Ishigaki Island, Okinawa. Here, we summarize the prevalence, Screening Library supplier incidence and spontaneous regression of AD and the relation of AD to other allergic diseases from previous reports. We also refer

to our recent findings from a population cohort study on Ishigaki Island, Okinawa.”
“There is great interest in molecules capable of inhibiting the interactions between p53 and its negative regulators hDM2 and hDMX, as these molecules have validated potential against cancers in which one or both oncoproteins are overexpressed. We reported previously that appropriately substituted beta(3)-peptides

inhibit these interactions and, more recently, that minimally cationic beta(3)-peptides are sufficiently cell permeable to upregulate p53-dependent genes in live cells. These observations, coupled with the known stability of beta-peptides in a cellular environment, and the recently reported structures of hDM2 and hDMX, motivated us to exploit computational modeling to identify beta-peptides with improved potency and/or selectivity. This exercise AG-881 cost successfully identified a new beta(3)-peptide, beta 53-16, that possesses the highly desirable attribute of high affinity for both hDM2 and hDMX and identifies the 3,4-dichlorophenyl moiety as a novel determinant of hDMX affinity.”
“Tension pneumocephalus complicating ventriculoperitoneal shunt is extremely rare. We report an elderly male who developed delayed tension pneumocephalus 12 months after ventriculoperitoneal shunt for hydrocephalus complicating aneurysmal subarachnoid hemorrhage. Fine-cut reformatted computer tomography scan revealed a large pneumatocele on the petrous apex associated with tegmen tympani defect. The shunt valve pressure was temporarily raised from 120 mm H(2)O to 200 mm H(2)O, and the patient underwent successful subtemporal extradural repair of the bony defect in the temporal bone.

Circ Res 98: 962-969, 2006) In this study, we targeted the vascu

Circ Res 98: 962-969, 2006). In this study, we targeted the vascular endothelium by using a lentivirus

construct expressing CYP4A2 under the control of the endothelium-specific promoter VE-cadherin (VECAD-4A2) and examined HKI272 the effect of long-term CYP4A2 overexpression on blood pressure and kidney function in SD rats. A bolus injection of VECAD-4A2 increased blood pressure (P < 0.001) by 26, 36, and 30 mmHg 10, 20, and 30 days postinjection, respectively. Arteries from VECAD-4A2-transduced rats produced increased levels of 20-HETE (P < 0.01), expressed lower levels of endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (p-eNOS) (P < 0.05), generated higher levels of superoxide anion, and displayed decreased relaxing responsiveness BI 2536 datasheet to acetylcholine (P < 0.05). Proteinuria increased by twofold in VECAD-4A2-transduced rats compared with controls. Treatment of VECAD-4A2-transduced rats with HET0016, an inhibitor of

20-HETE biosynthesis, not only attenuated the increase in blood pressure (P < 0.05) but also improved vascular function (acetylcholine-induced relaxations) and reduced plasma creatinine and proteinuria. HET0016 treatment decreased oxidative stress and increased the phosphorylated state of key proteins that regulate endothelial function, including eNOS, AKT, and AMPK. Collectively, these findings demonstrate that augmentation of vascular endothelial 20-HETE levels results in hypertension, endothelial dysfunction, and renal injury, which is offset by HET0016 through a reduction in vascular 20-HETE coupled with a lessening of oxidative stress and the amplification of pAKT, pAMPK, and p-eNOS levels leading

to normalization of endothelial responses.”
“Background-Vitamin D deficiency is highly prevalent and is associated with dyslipidemia and cardiovascular disease. VX-689 research buy The impact of correcting vitamin D deficiency on blood lipids, strong cardiovascular disease prognostic factors, is unknown.\n\nMethods and Results-To determine relationships between 25-hydroxyvitamin D levels and lipids, we analyzed 4.06 million deidentified patient laboratory test results from September 2009 through February 2011. We performed a cross-sectional study of this population to determine associations between 25-hydroxyvitamin D levels and lipids across clinically defined strata. We also conducted a retrospective cohort study of vitamin D deficient patients to investigate how changes in 25-hydroxyvitamin D levels relate to changes in lipid levels. After exclusions, 107 811 patients with serial testing were selected for cross-sectional analysis. Compared with vitamin D deficient patients (<20 ng/mL), those with optimal levels (>= 30 ng/mL) had lower mean total cholesterol (-1.

This paper summarizes the scientific basis, explains the diagnost

This paper summarizes the scientific basis, explains the diagnostic rationale and proves the concept by analyzing 5669 samples, where GFR and proteinuria work-up were

available. 63% (1446 of 2287) of the samples with a GFR above 60 showed either glomerular (37.8%, n = 865) or tubular proteinuria (25.4%, n = 581). The quantity of proteinuria increased severely with decreasing kidney function. The rate of glomerular proteinuria remained nearly constant in the different GFR groups, while primarily learn more tubular proteinuria increased from 23% to 63%. A proteinuria pattern indicating a good response to therapy was frequently combined with a high GFR (selective glomerular proteinuria/ incomplete tubular proteinuria), while the severe forms of unselective or complete tubular proteinuria associated with a severe

GFR decrease. Regression analysis showed a better inverse correlation of GFR with tubular (r = -0.643) than glomerular markers (r = -0.360; combined r = -0.646). We believe that this complex interrelated laboratory information must be delivered most effectively, i.e. with the use of a knowledge based system in combination with improved, visual oriented laboratory output. (C) 2009 European Federation of Internal Medicine. Published by Elsevier B.V. All rights reserved.”
“Aims: To assess visual outcome, tumour control and Emricasan chemical structure treatment-related morbidity in patients with optic nerve sheath meningiomas (ONSMs) treated with fractionated stereotactic radiotherapy (FSRT).\n\nPatients and methods: A retrospective

analysis of 45 patients (13 men and 32 women, median age 46 years) with ONSMs (51 optic nerves involved) treated in a single institution between 1997 and 2010 was carried out. FSRT was delivered to a dose of 50 Gy in 30 or 33 fractions as primary treatment in 39 patients and after surgery in six patients.\n\nResults: At a median follow-up of 30 months (range 1-13 years), the tumour control in 41 evaluable patients (four were lost to follow-up) was 100% at 5 years with no subsequent Selleck GDC-941 local or distant recurrence. Of the 46 evaluable optic nerves treated, 41 had residual vision (38 with impaired vision) before radiotherapy and five were blind in one eye. There was no recovery of vision in any of the blind eyes. Of 41 optic nerves with residual vision, 13 had improvement, 24 remained stable and four deteriorated; two patients (4%) developed radiation retinopathy. One patient developed a central retinal artery occlusion in the untreated eye 10 years after treatment.\n\nConclusion: FSRT is highly effective at controlling the growth of ONSMs with improvement or stabilisation of visual deficit in 89% of the optic nerves retaining some vision, albeit with a small risk of radiation-induced retinopathy. The results support the use of FSRT as an effective approach in the management of ONSM.

Upon coexpression of a modified Cloacin DF13 bacteriocin release

Upon coexpression of a modified Cloacin DF13 bacteriocin release protein, the hybrid proteins are released into the culture medium. This essentially applies to a distinguished reporter molecule, the green fluorescent protein, for which an extracellular production was not reported

so far. The sequestered proteins can be purified to approximate homogeneity by a simple, rapid and cheap procedure which utilizes the affinity of the maltose binding protein to alpha-1,4-glucans. (C) 2009 Elsevier B.V. All rights reserved.”
“In this review, we describe a new model to explain the regulation of myometrial function during pregnancy https://www.selleckchem.com/products/loxo-101.html and labour. We propose that the myometrium undergoes dramatic changes PX-478 manufacturer in phenotype from early pregnancy until the onset of labour, characterized by an early proliferative

phase, an intermediate phase of cellular hypertrophy and matrix elaboration, a third phase in which the cells assume a contractile phenotype and the final phase in which cells become highly active and committed to labour. The last phase of myometrial differentiation is Postpartum uterine involution, completing the reproductive cycle following pregnancy and labour by returning the uterus to its non-pregnant receptive state. We further propose that phenotypic modulation of the uterine myocytes is the result of integration of endocrine signals and mechanical stimulation of the uterus by the growing fetus. Our previous studies have shown that these signals are important in regulating the onset of labour and we now have indications that they regulate earlier myometrial smooth muscle differentiation.

We show that the high rate of myometrial cell proliferation in early pregnancy which reflects important aspects of many smooth muscle populations during development. The proliferative phenotype was associated with dramatic changes in the expression of IGF family proteins and coincided with an up-regulation of the anti-apoptotic pathway. Preliminary evidence suggests that myometrial hyperplasia was controlled by the PI3K-Akt-mTOR signaling pathway. The modulation of the mTOR pathway by rapamycin blocked the proliferative activity of the uterine myocytes. The growth and remodeling of the myometrium during pregnancy was associated with increased synthesis of extra cellular matrix (ECM) check details proteins and their corresponding integrin receptors. our results show a decrease in expression of fibrillar collagens and a coordinated temporal increase in expression of components of the basement membrane near term associated with decreased progesterone levels and increased mechanical tension. The phenotypic modulation of uterine smooth muscle cells during pregnancy culminates at term when a myometrium-specific conversion commits these cells to the labour phenotype, characterized by increased excitability, spontaneous activity, responsiveness to agonists and effective coupling of the myocytes.

Several works have demonstrated that lithium can either inhibit o

Several works have demonstrated that lithium can either inhibit or stimulate growth of normal and cancer cells. Hence, the present study is focused to analyze the underlying mechanisms that dictate the biphasic oncogenic properties of LiCl. In the current study, we have investigated the dose-dependent selleck effects of LiCl on human breast cancer cells (MCF-7) by assessing the consequences on cytotoxicity and protein expressions of signaling molecules crucial for the maintenance of cell survival. The results showed breast cancer cells respond in a diverse manner to LiCl, i.e., at lower concentrations (1, 5, and 10 mM), LiCl induces cell

survival by inhibiting apoptosis through regulation of GSK-3 beta, caspase-2, Bax, and cleaved caspase-7 and by activating anti-apoptotic proteins (Akt, beta-catenin, Bcl-2, and cyclin D1). In contrast, at high concentrations (50 and 100 mM), it induces apoptosis by reversing these effects. Moreover, LiCl also alters the sodium and potassium levels thereby altering the membrane potential of MCF-7 cells. Thus it is inferred that LiCl exerts a dose-dependent biphasic effect on breast cancer cells (MCF-7) by altering the apoptotic/anti-apoptotic balance.”
“Aminopeptidase N (APN/CD13) is one of the essential proteins for tumour invasion, angiogenesis and metastasis as it is over-expressed on the surface of different tumour cells. Based on our previous work that L-isoserine dipeptide derivatives

were potent APN inhibitors, we designed and synthesized L-isoserine tripeptide derivatives as APN inhibitors. Among these compounds, Screening Library cell line one compound 16l (IC50 = 2.51 +/- 0.2 +/- mu M) showed similar inhibitory effect compared with control compound Bestatin (IC50 = 6.25 +/- 0.4 mu M) and it could be used as novel lead compound for the APN inhibitors development as anticancer agents in the future.”
“The mechanisms governing the development of cardiac pacemaking and conduction system are not well understood. In order to provide evidence for AZD6244 chemical structure the derivation of pacemaking cells and the signal that induce and maintain the cells in the developing heart, Nkx2.5(+) cardiac progenitor cells (CPCs) were isolated from embryonic heart tubes of rats. Endothelin-1 was

subsequently added to the CPCs to induce differentiation of them towards cardiac pacemaking cells. After the treatment, Nkx2.5(+) CPCs displayed spontaneous beating and spontaneously electrical activity as what we have previously described. Furthermore, RT-PCR and immunofluorescence staining demonstrated that Tbx3 expression was increased and Nkx2.5 expression was decreased in the induced cells 4 days after ET-1 treatment. And the significantly increased expression of Hcn4 and connexin-45 were detected in the induced cells 10 days after the treatment. In addition, Nkx2.5(+) CPCs were transfected with pGCsi-Tbx3 4 days after ET-1 treatment in an attempt to determine the transcription regulatory factor governing the differentiation of the cells into cardiac pacemaking cells.


“Background: The gE protein of duck plague virus is the im


“Background: The gE protein of duck plague virus is the important membrane glycoprotein, its protein characterization has not been reported. In this study, we expressed and presented the characterization of the DPV gE product.\n\nResults: According to the sequence of the gE gene, a pair of primers were designed, and the DNA product with 1490bp in size was amplified by using the polymerase chain reaction (PCR). The PCR product was cloned learn more into pMD18-T vector, and subcloned into pET32a(+), generating the recombinant plasmid pET32a/DPV-gE. SDS-PAGE analysis showed that the fusion pET32a/DPV-gE protein was highly expressed after induction by 0.2 mM IPTG at 30 degrees C for 4.5 h in Rosseta

host cells. Over expressed 6xHis-gE fusion protein was purified by nickel affinity chromatography, and used to immunize the rabbits for the preparation of polyclonal antibody. The result of the intracellular localization revealed that the gE protein was appeared to be in the cytoplasm region. The real time PCR, RT-PCR analysis and Western Doramapimod solubility dmso blotting revealed that the gE gene was produced most abundantly during the late phase of replication in DPV-infected cells.\n\nConclusions: In this work, the DPV gE protein was successfully expressed in a prokaryotic expression system, and we presented the basic properties of the DPV gE

product for the first time. These properties of the gE protein provided a prerequisite for further functional analysis of this gene.”
“The therapy of ITP has recently been revolutionized with the introduction of thrombopoeitin stimulating agents. However, these medications are known to increase the platelet count only while the medication is being administered,

with SIS3 concentration a rapid fall of the platelet count to baseline pre-therapy levels on discontinuation. We report the case of a patient with chronic refractory ITP who has attained a prolonged remission after a short course of eltrombopag, with normalization of the platelet count, which is sustained 8 months after discontinuation of the medication.”
“Cell migration through tight interstitial spaces in three dimensional (3D) environments impacts development, wound healing and cancer metastasis and is altered by the aging process. The stiffness of the extracellular matrix (ECM) increases with aging and affects the cells and cytoskeletal processes involved in cell migration. However, the nucleus, which is the largest and densest organelle, has not been widely studied during cell migration through the ECM. Additionally, the nucleus is stiffened during the aging process through the accumulation of a mutant nucleoskeleton protein lamin A, progerin. By using microfabricated substrates to mimic the confined environment of surrounding tissues, we characterized nuclear movements and deformation during cell migration into micropillars where interspacing can be tuned to vary nuclear confinement.

The behavioral determinants of overweight differed between men an

The behavioral determinants of overweight differed between men and women, thus indicating the need for diverse gender-specific strategies.”
“This paper reviews the most common methods of generation of plasmas using microwaves with special emphasis on recently developed microwave plasma (MWP) sources for analytical applications. The art and science of microwave plasma optical and mass spectroscopy instrumentation (MWP-OES/MS), and the applications are briefly presented, including very recent advances Copanlisib cell line in the field as of 2012. The design and operation of MWPs is discussed to provide a basic understanding of the most important

selection criteria when designing MWP systems. The various plasma generation systems described include single-electrode capacitive microwave plasmas, electrodeless inductively coupled plasmas, multi-electrode systems energized with stationary or rotating fields. We also

discuss various technical realizations of MWP sources for selected applications. Examples of technical realizations of plasmas in closed structures (cavities), in open structures (surfatrons, planar plasma sources), and in magnetic fields (Hammer cavity) are discussed in detail. Finally, we mention micro-and mini-discharges as convenient sources for miniaturized spectrometric AZD9291 research buy systems. Specific topics include fundamental aspects of MWP, i.e., recent advances in the construction of analytical MK-1775 manufacturer MWPs (coaxially coupled cavities, strip-line technology, multi-point energizing, power combining, rotating field-excited plasmas), operational characteristics, analytical characteristics and applications. Special reference is made

to coupling with OES for determination of chromatographic effluents and particle sizing. The developments in elemental and molecular MS applications in both low-power and high-power MWPs are also discussed.”
“With the rise of high-throughput sequencing technology, traditional genotyping arrays are gradually being replaced by sequencing technology. Against this trend, Illumina has introduced an exome genotyping array that provides an alternative approach to sequencing, especially suited to large-scale genome-wide association studies (GWASs). The exome genotyping array targets the exome plus rare single-nucleotide polymorphisms (SNPs), a feature that makes it substantially more challenging to process than previous genotyping arrays that targeted common SNPs. Researchers have struggled to generate a reliable protocol for processing exome genotyping array data. The Vanderbilt Epidemiology Center, in cooperation with Vanderbilt Technologies for Advanced Genomics Analysis and Research Design (VANGARD), has developed a thorough exome chip-processing protocol. The protocol was developed during the processing of several large exome genotyping array-based studies, which included over 60,000 participants combined.